RNAi. Martin Latterich

Save this PDF as:
Size: px
Start display at page:

Download "RNAi. Martin Latterich"

Transcription

1 RNAi Martin Latterich

2

3 Contributors Abbreviations Preface i x ( xi xiii 1 Methods in RNA interference 1 Martin Latterich and Dalia Halawan i References 2 2 RNAi reagent design 3 Bernd Jagla and Nathalie Aulner 2.1 Introduction Lessons learned from X-ray structures/mechanism Current design considerations Asymmetry Sequence positional preferences Practical features Non-sequence position-based considerations shrna design considerations Prediction tools Other related tools Databases sirna databases mirna databases Target RNA secondary structure predictions Other resources on the web Concluding remarks 1 5 Acknowledgments 1 6 References RNAi - a chemical perspective 2 1 Ouathek Ouerfell i 3.1 Background Introduction sirnas versus shrnas sirnas shrnas RNAi reagents RNA chemistry 24

4 3.6 RNA synthesis methods High-throughput sirna synthesis: the full process Summary and outlook 2 7 Acknowledgments 2 8 References 28 4 Validation of RNAi 3 1 Nathalie Aulner and Bernd Jagla 4.1 Introduction sirna delivery sirna transfection Introduction of shrnas into mammalian cells RNAi screening delivery systems Silencing efficacy (potency) Detection of mrna levels Detection of protein levels Detection of knockdown efficiency using a reporter system (surrogate assays) Silencing validation sirna specificity Minimizing cell defense mechanism (dsrna interferon response) Conclusion 4 1 Acknowledgments 42 References RNAi libraries in dissecting molecular pathways of the human cell 47 Cheryl Eifert, Antonis Kourtidis and Douglas S. Conkli n 5.1 Introduction RNAi Approaches for loss-of-function screens High-throughput RNAi screens RNAi-induced phenotype selections Screens for mirna functions Perspectives in disease treatment 5 7 References High-throughput RNAi in Caenorhabditis elegans - from molecular phenotypes to pathway analysis 6 5 Sarah Jenna and Eric Cheve t 6.1 Introduction RNAi in C. elegans High-throughput RNAi in C. elegans The experiments Summary 68 References 69 Protocol 6.1 : Generation of constructs driving RNAi through a feeding procedure 7 1 Protocol 6.2: RNAi treatment of GFP reporter animals 73 Protocol 6.3 : Sorting of fluorescent animals and measurement of the UPR 77

5 7 RNAi in Xenopus laevis 79 Adrianna L. Stromme and Craig A. Mandato 7.1 Introduction Oocyte isolation Inducing ovulation Collecting eggs Testes isolation In vitro fertilization Microinjecting dsrna into embryos/oocytes Dejellying embryos Vitelline membrane removal Microinjections Lineage labeling Dextran amines ß-Galactosidase RNA GFP RNA as a lineage marker Screening of phenotypes 8 5 References 8 5 Protocol 7.1 : Solutions appendix 8 6 Protocol 7.2: X-gal staining protocol (Sive et al., 1997) 8 8 Protocol 7.3: Overall protocol for sirna experiment (example) Generation of transgenic and knockdown mice with lentiviral vectors and RNAi techniques 9 1 Jenni Huusko, Petri I. Mäkinen, Leena Alhonen and Seppo Ylä-Herttual a 8.1 Introduction Production of transgenic and knockdown mice Use of ES cells Use of embryos Lentivirus vectors Design of LVs for the generation of knockdown mice Constitutive pol III promoters Regulatable pol III promoters Pol II promoters 9 5 References 9 6 Protocol 8.1: Mice, reagents and equipment 9 9 Protocol 8.2: Setting up capillaries, injection needles, injection chambers an d preparations for transgenesis 10 2 Protocol 8.3: Direct microinjection of the viral construct to the subzonal spac e (= perivitelline space) of a fertilized egg cell 10 5 Protocol 8.4: Zona pellucida removal and lentiviral transduction RNAi in fungi 11 3 Hitoshi Nakayashiki 9.1 Introduction The discovery of quelling in Neurospora Meiotic silencing by unpaired DNA (MSUD), a nove l gene-silencing phenomenon in Neurospora RNAi as a genetic tool in fungi 114

6 9.2 RNAi strategies in fungi RNAi using a hairpin RNA-expressing plasmid RNAi using an opposing-dual promoter system Direct delivery of dsrna into fungal cells Simultaneous silencing of multiple genes Genetic transformation and RNAi protocols for fungi 119 Acknowledgments 12 0 References 120 Protocol 9.1 : Transformation of Magnaporthe oryzae by the calcium chloride/polyethylene glycol (PEG) method 12 3 Protocol 9.2: Transformation of Cryptococcus neoformans by electroporation 12 5 Protocol 9.3: Transformation of Mortierella alpina by the microparticl e bombardment method 12 7 Protocol 9.4: Transformation of Phytophthora infestans by th e Lipofectin-mediated transfection method 12 9 Index 133

RNAi Shooting the Messenger!

RNAi Shooting the Messenger! RNAi Shooting the Messenger! Bronya Keats, Ph.D. Department of Genetics Louisiana State University Health Sciences Center New Orleans Email: bkeats@lsuhsc.edu RNA interference (RNAi) A mechanism by which

More information

Protein Expression. A Practical Approach J. HIGGIN S

Protein Expression. A Practical Approach J. HIGGIN S Protein Expression A Practical Approach S. J. HIGGIN S B. D. HAMES List of contributors Abbreviations xv Xvi i 1. Protein expression in mammalian cell s Marlies Otter-Nilsson and Tommy Nilsso n 1. Introduction

More information

岑 祥 股 份 有 限 公 司 技 術 專 員 費 軫 尹 20100803

岑 祥 股 份 有 限 公 司 技 術 專 員 費 軫 尹 20100803 技 術 專 員 費 軫 尹 20100803 Overview of presentation Basic Biology of RNA interference Application of sirna for gene function? How to study mirna? How to deliver sirna and mirna? New prospects on RNAi research

More information

Lezioni Dipartimento di Oncologia Farmacologia Molecolare. RNA interference. Giovanna Damia 29 maggio 2006

Lezioni Dipartimento di Oncologia Farmacologia Molecolare. RNA interference. Giovanna Damia 29 maggio 2006 Lezioni Dipartimento di Oncologia Farmacologia Molecolare RNA interference Giovanna Damia 29 maggio 2006 RNA INTERFERENCE Sequence-specific gene suppression by dsrnas Gene silencing by dsrna: C. elegans

More information

TOOLS sirna and mirna. User guide

TOOLS sirna and mirna. User guide TOOLS sirna and mirna User guide Introduction RNA interference (RNAi) is a powerful tool for suppression gene expression by causing the destruction of specific mrna molecules. Small Interfering RNAs (sirnas)

More information

EXPRESSION ARREST shrna mir GENOME- WIDE LIBRARIES

EXPRESSION ARREST shrna mir GENOME- WIDE LIBRARIES C GUGAAG EXPRESSION ARREST shrna mir GENOME- WIDE LIBRARIES MicroRNA-adapted shrna (shrna mir ) for increased, specific and consistent knockdown. MicroRNA PROCESSING PATHWAY UTILIZED FOR shrna mir Developed

More information

Transfection-Transfer of non-viral genetic material into eukaryotic cells. Infection/ Transduction- Transfer of viral genetic material into cells.

Transfection-Transfer of non-viral genetic material into eukaryotic cells. Infection/ Transduction- Transfer of viral genetic material into cells. Transfection Key words: Transient transfection, Stable transfection, transfection methods, vector, plasmid, origin of replication, reporter gene/ protein, cloning site, promoter and enhancer, signal peptide,

More information

Biosafety considerations of new dsrna molecules

Biosafety considerations of new dsrna molecules Biosafety considerations of new dsrna molecules GM-free Europe Conference 2015 May 6-8 Berlin, Germany Dr. Sarah Agapito Current GMOs Plant transformed to produce new protein A G C U G G C A A U G U C

More information

Recombinant DNA and Biotechnology

Recombinant DNA and Biotechnology Recombinant DNA and Biotechnology Chapter 18 Lecture Objectives What Is Recombinant DNA? How Are New Genes Inserted into Cells? What Sources of DNA Are Used in Cloning? What Other Tools Are Used to Study

More information

Product: Expression Arrest TM egfp control shrna vector

Product: Expression Arrest TM egfp control shrna vector Product: Expression Arrest TM egfp control vector Catalog #: RHS1702 Product Description The laboratory of Dr. Greg Hannon at Cold Spring Harbor Laboratory (CSHL) has created an RNAi Clone Library comprised

More information

2.1.2 Characterization of antiviral effect of cytokine expression on HBV replication in transduced mouse hepatocytes line

2.1.2 Characterization of antiviral effect of cytokine expression on HBV replication in transduced mouse hepatocytes line i 1 INTRODUCTION 1.1 Human Hepatitis B virus (HBV) 1 1.1.1 Pathogenesis of Hepatitis B 1 1.1.2 Genome organization of HBV 3 1.1.3 Structure of HBV virion 5 1.1.4 HBV life cycle 5 1.1.5 Experimental models

More information

Dicer Substrate RNAi Design

Dicer Substrate RNAi Design INTEGRATED DNA TECHNOLOGIES, INC. Dicer Substrate RNAi Design How to design and order 27-mer Dicer-substrate Duplex RNAs for use as RNA interference reagents The following document provides a summary of

More information

Functional and Biomedical Aspects of Genome Research

Functional and Biomedical Aspects of Genome Research Functional and Biomedical Aspects of Genome Research 20 11 35 Vorlesung SS 04 Bartsch, Jockusch & Schmitt-John Mi. 9:15-10:00, in W7-135 13 Functional RNAs Thomas Schmitt-John micro RNAs small interfering

More information

CODICE DESCRIZIONE QUANTITA' PREZZO EURO

CODICE DESCRIZIONE QUANTITA' PREZZO EURO Broad Spectrum Plasmid DNA and sirna/mirna Transfection MIR 6000 TransIT-X2 Dynamic Delivery System 1.5 ml 823,35 MIR 6003 TransIT-X2 Dynamic Delivery System 0.3 ml 196,35 MIR 6004 TransIT-X2 Dynamic Delivery

More information

2012 CCR Mouse Research Models. Laboratory Animal Sciences Program CORE Resources

2012 CCR Mouse Research Models. Laboratory Animal Sciences Program CORE Resources National Cancer Institute 2012 CCR Mouse Research Models Mouse 101 Laboratory Animal Sciences Program CORE Resources Lionel Feigenbaum, Ph.D. Laboratory Animal Sciences Program May 7, 2012 CORE Resources

More information

Von Mäusen und Menschen E - 1

Von Mäusen und Menschen E - 1 Von Mäusen und Menschen E - 1 Mus musculus: Genetic Portrait of the House Mouse E - 3 Outline Mouse genome Mouse life cycle Transgenic protocols Addition of genes by nuclear injection Removal of genes

More information

MicroRNA formation. 4th International Symposium on Non-Surgical Contraceptive Methods of Pet Population Control

MicroRNA formation. 4th International Symposium on Non-Surgical Contraceptive Methods of Pet Population Control MicroRNA formation mirna s are processed from several precursor stages Mammalian genomes seem to have 100 s of mirna s Nucleotides in positions 2-8 of an mirna are considered the mirna seed 5 Methyl-G

More information

Dicer-Substrate sirna Technology

Dicer-Substrate sirna Technology As featured in BioRadiations 120 Dicer-Substrate sirna Technology Advances in sirna Designs Improve Gene-Specific Silencing Steve Kulisch, Teresa Esch, Christina Whitman-Guliaev, and Teresa Rubio, Bio-Rad

More information

Understanding the immune response to bacterial infections

Understanding the immune response to bacterial infections Understanding the immune response to bacterial infections A Ph.D. (SCIENCE) DISSERTATION SUBMITTED TO JADAVPUR UNIVERSITY SUSHIL KUMAR PATHAK DEPARTMENT OF CHEMISTRY BOSE INSTITUTE 2008 CONTENTS Page SUMMARY

More information

How to construct transgenic mice

How to construct transgenic mice How to construct transgenic mice Sandra Beer-Hammer Autumn School 2011 Bad Schandau Pharmakologie und Experimentelle Therapie (APET) Overview History Generation of embryonic stem (ES) cell lines Generation

More information

Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College

Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Primary Source for figures and content: Eastern Campus Tortora, G.J. Microbiology

More information

An Overview of Cells and Cell Research

An Overview of Cells and Cell Research An Overview of Cells and Cell Research 1 An Overview of Cells and Cell Research Chapter Outline Model Species and Cell types Cell components Tools of Cell Biology Model Species E. Coli: simplest organism

More information

The world of non-coding RNA. Espen Enerly

The world of non-coding RNA. Espen Enerly The world of non-coding RNA Espen Enerly ncrna in general Different groups Small RNAs Outline mirnas and sirnas Speculations Common for all ncrna Per def.: never translated Not spurious transcripts Always/often

More information

Recombinant DNA Technology

Recombinant DNA Technology Recombinant DNA Technology Dates in the Development of Gene Cloning: 1965 - plasmids 1967 - ligase 1970 - restriction endonucleases 1972 - first experiments in gene splicing 1974 - worldwide moratorium

More information

CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA

CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA INTRODUCTION DNA : DNA is deoxyribose nucleic acid. It is made up of a base consisting of sugar, phosphate and one nitrogen base.the

More information

Especially developed for the transfection of mammalian cells with sirna or mirna

Especially developed for the transfection of mammalian cells with sirna or mirna METAFECTENE SI Especially developed for the transfection of mammalian cells with sirna or mirna For ordering information, MSDS, publications and application notes see www.biontex.com Description Cat. No.

More information

GeneCopoeia Genome Editing Tools for Safe Harbor Integration in. Mice and Humans. Ed Davis, Liuqing Qian, Ruiqing li, Junsheng Zhou, and Jinkuo Zhang

GeneCopoeia Genome Editing Tools for Safe Harbor Integration in. Mice and Humans. Ed Davis, Liuqing Qian, Ruiqing li, Junsheng Zhou, and Jinkuo Zhang G e n e C o p o eia TM Expressway to Discovery APPLICATION NOTE Introduction GeneCopoeia Genome Editing Tools for Safe Harbor Integration in Mice and Humans Ed Davis, Liuqing Qian, Ruiqing li, Junsheng

More information

An Introduction to Transfection Methods Technical Reference Guide

An Introduction to Transfection Methods Technical Reference Guide An Introduction to Transfection Methods Transfection Technologies The introduction of nucleic acids into cells is one of the most valuable and frequently used tools of biological science. Transfection

More information

Outline. interfering RNA - What is dat? Brief history of RNA interference. What does it do? How does it work?

Outline. interfering RNA - What is dat? Brief history of RNA interference. What does it do? How does it work? Outline Outline interfering RNA - What is dat? Brief history of RNA interference. What does it do? How does it work? What is RNA interference? Recently discovered regulatory level. Genome immune system.

More information

LONDON REGIONAL TRANSGENIC AND GENE TARGETING FACILITY

LONDON REGIONAL TRANSGENIC AND GENE TARGETING FACILITY LONDON REGIONAL TRANSGENIC AND GENE TARGETING FACILITY SPERM CRYOPRESERVATION AND IN VITRO FERTILIZATION RESOURCE PACKAGE For information, contact: Dr. Christopher Pin, PhD, Scientific Director London

More information

Zebrafish Chemical Screens

Zebrafish Chemical Screens Zebrafish Chemical Screens 1 Historical look at zebrafish chemical screens 2 The effects of AAF on embryonic development in relation to its carcinogenic role 3 First generation Screens HISAOKA, K. K. (1958).

More information

Manual for: sirna-trans Maximo Reagent

Manual for: sirna-trans Maximo Reagent Manual for: sirna-trans Maximo Reagent Contact: info@geneon.net; www.geneon.net GeneON GmbH Germany GeneON GmbH / Protocol sirna-trans reagent vs. 1.3 / www.geneon.net / - 0 - sirna-trans reagent Instruction

More information

Engineering of Yellow Mosaic Virus Resistance (YMVR) in Blackgram. Project ID: 1 April 2000 to 31 August 2004. Project Duration:

Engineering of Yellow Mosaic Virus Resistance (YMVR) in Blackgram. Project ID: 1 April 2000 to 31 August 2004. Project Duration: Engineering of Yellow Mosaic Virus Resistance (YMVR) in Blackgram ID: Duration: Coordinator in Switzerland: PS1 1 April 2000 to 31 August 2004 Prof. Thomas Hohn University of Basel Botanisches Institut

More information

sirna Duplexes & RNAi Explorer

sirna Duplexes & RNAi Explorer P r o d u c t S p e c i f i c a t i o n s Guaranteed RNAi Explorer kit with Fl/Dabcyl Molecular Beacon Catalog No.:27-6402-01 Guaranteed RNAi Explorer kit with Fluorescein/Tamra TaqMan Catalog No.:27-6402-01

More information

Notch 1 -dependent regulation of cell fate in colorectal cancer

Notch 1 -dependent regulation of cell fate in colorectal cancer Notch 1 -dependent regulation of cell fate in colorectal cancer Referees: PD Dr. Tobias Dick Prof. Dr. Wilfried Roth http://d-nb.info/1057851272 CONTENTS Summary 1 Zusammenfassung 2 1 INTRODUCTION 3 1.1

More information

How to construct transgenic mice

How to construct transgenic mice How to construct transgenic mice Sandra Beer-Hammer Autumn School 2012 Bad Schandau Pharmakologie und Experimentelle Therapie (APET) Overview History Generation of embryonic stem (ES) cell lines Generation

More information

RNA Viruses. A Practical Approac h. Alan J. Cann

RNA Viruses. A Practical Approac h. Alan J. Cann RNA Viruses A Practical Approac h Alan J. Cann List of protocols page xiii Abbreviations xvii Investigation of RNA virus genome structure 1 A j. Easton, A.C. Marriott and C.R. Pringl e 1 Introduction-the

More information

Instructions. Torpedo sirna. Material. Important Guidelines. Specifications. Quality Control

Instructions. Torpedo sirna. Material. Important Guidelines. Specifications. Quality Control is a is a state of the art transfection reagent, specifically designed for the transfer of sirna and mirna into a variety of eukaryotic cell types. is a state of the art transfection reagent, specifically

More information

How to construct transgenic mice

How to construct transgenic mice How to construct transgenic mice Sandra Beer-Hammer Autumn School 2010 Bad Schandau Methods additional genetic information transgenic mouse line gene inactivation gene-deficient knockout mouse line Jak2

More information

Trasposable elements: P elements

Trasposable elements: P elements Trasposable elements: P elements In 1938 Marcus Rhodes provided the first genetic description of an unstable mutation, an allele of a gene required for the production of pigment in maize. This instability

More information

Machine Learning Approaches to sirna Efficacy Prediction

Machine Learning Approaches to sirna Efficacy Prediction Machine Learning Approaches to sirna Efficacy Prediction by Sahar Abubucker B.E., Madras University, 2000 THESIS Submitted in Partial Fulfillment of the Requirements for the Degree of Master of Science

More information

Cell Discovery 360: Explore more possibilities.

Cell Discovery 360: Explore more possibilities. Cell Discovery 360: Explore more possibilities. Flexible solutions for the BioPharma laboratory Blood Banking Capillary Electrophoresis Centrifugation Flow Cytometry Genomics Lab Automation Lab Tools Particle

More information

QIAGEN Transfection Technologies

QIAGEN Transfection Technologies QIAGEN Transfection Technologies Efficient and robust transfection for all your applications Sample & Assay Technologies Table of contents QIAGEN solutions for efficient transfection Transfection is a

More information

Inhibition of MicroRNA Sensitizes 3D Breast Cancer Microtissues to Radiation Therapy

Inhibition of MicroRNA Sensitizes 3D Breast Cancer Microtissues to Radiation Therapy CASE STUDY High Content Image Analysis Inhibition of MicroRNA Sensitizes 3D Breast Cancer Microtissues to Radiation Therapy Dr. Nataša Anastasov leads the Personalized Radiation Therapy Group within the

More information

Bacterial Transformation with Green Fluorescent Protein. Table of Contents Fall 2012

Bacterial Transformation with Green Fluorescent Protein. Table of Contents Fall 2012 Bacterial Transformation with Green Fluorescent Protein pglo Version Table of Contents Bacterial Transformation Introduction..1 Laboratory Exercise...3 Important Laboratory Practices 3 Protocol...... 4

More information

QUICK CELL CAPTURE AND CHARACTERIZATION GUIDE FOR CELLSEARCH CUSTOMERS

QUICK CELL CAPTURE AND CHARACTERIZATION GUIDE FOR CELLSEARCH CUSTOMERS QUICK CELL CAPTURE AND CHARACTERIZATION GUIDE FOR CELLSEARCH CUSTOMERS CellSave EDTA Blood sample Rare cell capture Enumeration Single protein marker Cell capture for molecular characterization CELLSEARCH

More information

SureSilencing shrna Plasmids

SureSilencing shrna Plasmids User Manual BIOMOL GmbH Waidmannstr. 35 22769 Hamburg info@biomol.de www.biomol.de Phone:+49-40-8532600 or 0800-2466651 (D) Fax: +49-40-85326022 or 0800-2466652 (D) SureSilencing shrna Plasmids Genome-Wide

More information

!!!!!!!!!!!!!!!!!!!!!!!!!!

!!!!!!!!!!!!!!!!!!!!!!!!!! Figure S7 cdl-1 3'UTR gld-1 binding site (4..10) let-7 binding site (326..346) Figure Legends Figure S1. gld-1 genetically interacts with nhl-2 and vig-1 during germline development. (A) nhl-2(ok818) enhances

More information

HiPerFect Transfection Reagent Handbook

HiPerFect Transfection Reagent Handbook Fifth Edition October 2010 HiPerFect Transfection Reagent Handbook For transfection of eukaryotic cells with sirna and mirna Sample & Assay Technologies QIAGEN Sample and Assay Technologies QIAGEN is the

More information

Guidance. 2. Definitions. 1. Introduction

Guidance. 2. Definitions. 1. Introduction Work with naked DNA or RNA (including oligonucleotides, sirna, mirna, sequences that code for highly biologically active molecules and full length viral genomes) Guidance 1. Introduction The following

More information

How many of you have checked out the web site on protein-dna interactions?

How many of you have checked out the web site on protein-dna interactions? How many of you have checked out the web site on protein-dna interactions? Example of an approximately 40,000 probe spotted oligo microarray with enlarged inset to show detail. Find and be ready to discuss

More information

Diabetes and Drug Development

Diabetes and Drug Development Diabetes and Drug Development Metabolic Disfunction Leads to Multiple Diseases Hypertension ( blood pressure) Metabolic Syndrome (Syndrome X) LDL HDL Lipoproteins Triglycerides FFA Hyperinsulinemia Insulin

More information

Primetime for KNIME:

Primetime for KNIME: Primetime for KNIME: Towards an Integrated Analysis and Visualization Environment for RNAi Screening Data F. Oliver Gathmann, Ph. D. Director IT, Cenix BioScience Presentation for: KNIME User Group Meeting

More information

Innate Immunity. Insects rely solely on an innate immune system for defense against infection

Innate Immunity. Insects rely solely on an innate immune system for defense against infection Innate Immunity Insects rely solely on an innate immune system for defense against infection The importance of mammalian innate immunity has only recently become appreciated The innate immune response

More information

Micro RNAs: potentielle Biomarker für das. Blutspenderscreening

Micro RNAs: potentielle Biomarker für das. Blutspenderscreening Micro RNAs: potentielle Biomarker für das Blutspenderscreening micrornas - Background Types of RNA -Coding: messenger RNA (mrna) -Non-coding (examples): Ribosomal RNA (rrna) Transfer RNA (trna) Small nuclear

More information

Mir-X mirna First-Strand Synthesis Kit User Manual

Mir-X mirna First-Strand Synthesis Kit User Manual User Manual Mir-X mirna First-Strand Synthesis Kit User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Clontech Laboratories, Inc.

More information

Thermo Scientific Dharmacon sirna Libraries

Thermo Scientific Dharmacon sirna Libraries Thermo Scientific Dharmacon sirna Libraries Scientific innovation in sirna potency, specificity and delivery Superior sirna in comprehensive collections for reliable screens Market leading products for

More information

Germ cell formation / gametogenesis And Fertilisation

Germ cell formation / gametogenesis And Fertilisation Developmental Biology BY1101 P. Murphy Lecture 3 The first steps to forming a new organism Descriptive embryology I Germ cell formation / gametogenesis And Fertilisation Why bother with sex? In terms of

More information

AP Biology Essential Knowledge Student Diagnostic

AP Biology Essential Knowledge Student Diagnostic AP Biology Essential Knowledge Student Diagnostic Background The Essential Knowledge statements provided in the AP Biology Curriculum Framework are scientific claims describing phenomenon occurring in

More information

Twincore - Zentrum für Experimentelle und Klinische Infektionsforschung Institut für Molekulare Bakteriologie

Twincore - Zentrum für Experimentelle und Klinische Infektionsforschung Institut für Molekulare Bakteriologie Twincore - Zentrum für Experimentelle und Klinische Infektionsforschung Institut für Molekulare Bakteriologie 0 HELMHOLTZ I ZENTRUM FÜR INFEKTIONSFORSCHUNG Technische Universität Braunschweig Institut

More information

Gene Therapy. The use of DNA as a drug. Edited by Gavin Brooks. BPharm, PhD, MRPharmS (PP) Pharmaceutical Press

Gene Therapy. The use of DNA as a drug. Edited by Gavin Brooks. BPharm, PhD, MRPharmS (PP) Pharmaceutical Press Gene Therapy The use of DNA as a drug Edited by Gavin Brooks BPharm, PhD, MRPharmS (PP) Pharmaceutical Press Contents Preface xiii Acknowledgements xv About the editor xvi Contributors xvii An introduction

More information

RNA Structure and folding

RNA Structure and folding RNA Structure and folding Overview: The main functional biomolecules in cells are polymers DNA, RNA and proteins For RNA and Proteins, the specific sequence of the polymer dictates its final structure

More information

Microarray Technology

Microarray Technology Microarrays And Functional Genomics CPSC265 Matt Hudson Microarray Technology Relatively young technology Usually used like a Northern blot can determine the amount of mrna for a particular gene Except

More information

BIOTECHNOLOGY OPERATIONS

BIOTECHNOLOGY OPERATIONS BIOTECHNOLOGY OPERATIONS Principles and Practices Michael J. Roy TECHNISCHE INFORMATION SBIBLIOTHEK UNIVERSITATSBIBLIOTHEK HANNOVER CRC Press TaylorStFrancis Croup Boca Raton London New York CRC Press

More information

CONTRACTING ORGANIZATION: University of Alabama at Birmingham Birmingham, AL 35294

CONTRACTING ORGANIZATION: University of Alabama at Birmingham Birmingham, AL 35294 AD Award Number: W81XWH-08-1-0030 TITLE: Regulation of Prostate Cancer Bone Metastasis by DKK1 PRINCIPAL INVESTIGATOR: Gregory A. Clines, M.D., Ph.D. CONTRACTING ORGANIZATION: University of Alabama at

More information

Chapter Five Development of reporter systems for probing the mirna pathway

Chapter Five Development of reporter systems for probing the mirna pathway Chapter Five Development of reporter systems for probing the mirna pathway 1. Introduction 1.1. Why study mirna biogenesis and effector pathways? Understanding mirna biogenesis and downstream effector

More information

ELITE Custom Antibody Services

ELITE Custom Antibody Services ELITE Custom Antibody Services ELITE Custom Antibody Services Experience, confidence, and understanding As a manufacturer and service provider, we have the experience, confidence, and understanding to

More information

1865 Discovery: Heredity Transmitted in Units

1865 Discovery: Heredity Transmitted in Units 1859 Discovery: Natural Selection Genetic Timeline Charles Darwin wrote On the Origin of Species by Means of Natural Selection, or the Preservation of Favored Races in the Struggle for Life. 1865 Discovery:

More information

Accelerating drug development to FTIH: Potential of new expression technologies

Accelerating drug development to FTIH: Potential of new expression technologies Accelerating drug development to FTIH: Potential of new expression technologies Lekan Daramola Associate Director Biopharmaceutical Development, Cell Culture & Fermentation Sciences CMC Strategy Forum

More information

Arabidopsis. A Practical Approach. Edited by ZOE A. WILSON Plant Science Division, School of Biological Sciences, University of Nottingham

Arabidopsis. A Practical Approach. Edited by ZOE A. WILSON Plant Science Division, School of Biological Sciences, University of Nottingham Arabidopsis A Practical Approach Edited by ZOE A. WILSON Plant Science Division, School of Biological Sciences, University of Nottingham OXPORD UNIVERSITY PRESS List of Contributors Abbreviations xv xvu

More information

OriGene Technologies, Inc. MicroRNA analysis: Detection, Perturbation, and Target Validation

OriGene Technologies, Inc. MicroRNA analysis: Detection, Perturbation, and Target Validation OriGene Technologies, Inc. MicroRNA analysis: Detection, Perturbation, and Target Validation -Optimal strategies to a successful mirna research project Optimal strategies to a successful mirna research

More information

Chapter 23 Definitions of GMO/LMO and modern biotechnology. Three different definitions but the same legal interpretation?

Chapter 23 Definitions of GMO/LMO and modern biotechnology. Three different definitions but the same legal interpretation? Chapter 23 Definitions of GMO/LMO and modern biotechnology JAN HUSBY NORWEGIAN INSTITUTE OF GENE ECOLOGY (GENØK), TROMSØ, NORWAY Three different definitions but the same legal interpretation? There are

More information

DNA Fingerprinting. Unless they are identical twins, individuals have unique DNA

DNA Fingerprinting. Unless they are identical twins, individuals have unique DNA DNA Fingerprinting Unless they are identical twins, individuals have unique DNA DNA fingerprinting The name used for the unambiguous identifying technique that takes advantage of differences in DNA sequence

More information

BIOPATENTS IN CHINA. Christopher Shaowei Heather Lin. April 4, 2014, Budapest, Hungary 2014/4/4 NTD IP ATTORNEYS

BIOPATENTS IN CHINA. Christopher Shaowei Heather Lin. April 4, 2014, Budapest, Hungary 2014/4/4 NTD IP ATTORNEYS BIOPATENTS IN CHINA Christopher Shaowei Heather Lin April 4, 2014, Budapest, Hungary Unpatentable subject matter Art 5, Patent Law: No patent right shall be granted for any invention-creation that is contrary

More information

The Leading Gene Through Screen CRO

The Leading Gene Through Screen CRO The Leading Gene Through Screen CRO Molecular Biology Services Molecular Biology drives modern drug discovery. Drug screening, target validation and structural biology processes frequently require recombinant

More information

LightSwitch Luciferase Assay System

LightSwitch Luciferase Assay System Constructs, Assays & Services Assay System Promoter GoClones Synthetic Response Elements Pathway Cell Lines 3 UTR GoClones mirna Mimics & Inhibitors Synthetic mirna Targets Transfection Reagents Assay

More information

Practical Cell Analysis

Practical Cell Analysis Practical Cell Analysis Dimitri Pappas Dept of Chemistry & Biochemistry, Texas Tech University, USA WILEY A John Wiley and Sons, Ltd, Publication Contents Preface Acknowledgments xiii xix 1 Getting Started

More information

Assisted Reproductive Technologies at IGO

Assisted Reproductive Technologies at IGO 9339 Genesee Avenue, Suite 220 San Diego, CA 92121 858 455 7520 Assisted Reproductive Technologies at IGO Although IGO no longer operates an IVF laboratory or program as such, we work closely with area

More information

Optimized Tandem amirna Mediates Stronger Inhibitory Effects on Hepatitis B Virus Infection

Optimized Tandem amirna Mediates Stronger Inhibitory Effects on Hepatitis B Virus Infection Optimized Tandem amirna Mediates Stronger Inhibitory Effects on Hepatitis B Virus Infection Chunwen Pu 1,2, Lifen Wang 2,*, Xiaohui Miao 3,*, Yong Zhang 1, Chunmeng Jiang 2, Wei Liu 1, Weixiang Sun 1,

More information

E-RNAi: a web application to design optimized RNAi constructs

E-RNAi: a web application to design optimized RNAi constructs W582 W588 Nucleic Acids Research, 2005, Vol. 33, Web Server issue doi:10.1093/nar/gki468 E-RNAi: a web application to design optimized RNAi constructs Zeynep Arziman, Thomas Horn and Michael Boutros* Boveri-Group

More information

BioMmune Technologies Inc. Corporate Presentation 2015

BioMmune Technologies Inc. Corporate Presentation 2015 BioMmune Technologies Inc Corporate Presentation 2015 * Harnessing the body s own immune system to fight cancer & other autoimmune diseases BioMmune Technologies Inc. (IMU) ABOUT A public biopharmaceutical

More information

GenScript sirna Cassette Protocol

GenScript sirna Cassette Protocol GenScript sirna Cassette Protocol Technical Manual No. 0168 Version 20040510 I Introduction.... 1 II sirna Cassette.... 1 III Product Description. 2 IV Transfecting Mammalian Cells. 3 V References... 5

More information

PRODUCT INFORMATION...

PRODUCT INFORMATION... VIROMER RED In vitro plasmid DNA and mrna Standard Transfection PRODUCT INFORMATION... 2 GENERAL... 2 RED OR YELLOW?... 3 PROTOCOL GUIDELINES... 4 GENERAL REMARKS... 4 CELL CULTURE AND PLATING... 4 FORWARD/REVERSE

More information

Embryonic & and induced pluripotent Stem Cells. May 2010 Dipl. Biol. Dr. Kurt Pfannkuche

Embryonic & and induced pluripotent Stem Cells. May 2010 Dipl. Biol. Dr. Kurt Pfannkuche Embryonic & and induced pluripotent Stem Cells May 2010 Dipl. Biol. Dr. Kurt Pfannkuche Seite 2 What makes a stem cell? open chromatin structure plasticity! Seite 3 self-renewal Plasticity Seite 4 Culture

More information

13.4 Gene Regulation and Expression

13.4 Gene Regulation and Expression 13.4 Gene Regulation and Expression Lesson Objectives Describe gene regulation in prokaryotes. Explain how most eukaryotic genes are regulated. Relate gene regulation to development in multicellular organisms.

More information

Supplemental Information. McBrayer et al. Supplemental Data

Supplemental Information. McBrayer et al. Supplemental Data 1 Supplemental Information McBrayer et al. Supplemental Data 2 Figure S1. Glucose consumption rates of MM cell lines exceed that of normal PBMC. (A) Normal PBMC isolated from three healthy donors were

More information

The Potential of RNA interference & The Future of DNA directed RNAi Therapeutics

The Potential of RNA interference & The Future of DNA directed RNAi Therapeutics The Potential of RNA interference & The Future of DNA directed RNAi Therapeutics November 2010 This presentation contains forward looking statements that involve risks and uncertainties. Although we believe

More information

Functional RNAs; RNA catalysts, mirna,

Functional RNAs; RNA catalysts, mirna, Functional RNAs; RNA catalysts, mirna, srna, RNAi... RNAs have many functions rrna (ribosomal RNA) trna (transfer RNA) mrna (Messenger RNA) snrna (including snorna) ) (Small nuclear RNA- splicing) Other

More information

The following chapter is called "Preimplantation Genetic Diagnosis (PGD)".

The following chapter is called Preimplantation Genetic Diagnosis (PGD). Slide 1 Welcome to chapter 9. The following chapter is called "Preimplantation Genetic Diagnosis (PGD)". The author is Dr. Maria Lalioti. Slide 2 The learning objectives of this chapter are: To learn the

More information

Andrew F. Roberts, Ph.D. Deputy Director, CERA. September 19, 2013

Andrew F. Roberts, Ph.D. Deputy Director, CERA. September 19, 2013 Risk assessment and regulation of RNAi Pesticides in the context of GE plants and the regulation of novel plant breeding techniques in the United States Andrew F. Roberts, Ph.D. Deputy Director, CERA September

More information

Post-transcriptional control of gene expression

Post-transcriptional control of gene expression Post-transcriptional control of gene expression Possible post-transcriptional controls on gene expression Only a few of these controls are likely to be important for any one gene Mecanismo de processamento

More information

MEF Starter Nucleofector Kit

MEF Starter Nucleofector Kit page 1 of 7 MEF Starter Nucleofector Kit for Mouse Embryonic Fibroblasts (MEF) MEF display significant phenotypic variations which depend on the strain, the genetic background of the mice they are isolated

More information

Animal Cell Culture. Third Edition. A Practical Approach OXJORD VNIVVRSITY 1'RVSS

Animal Cell Culture. Third Edition. A Practical Approach OXJORD VNIVVRSITY 1'RVSS Animal Cell Culture Third Edition A Practical Approach Edited by John R. W. Masters 3rd Floor Research Laboratories, University College London OXJORD VNIVVRSITY 1'RVSS Contents List of protocols page xiii

More information

BLOCK-iT Products: Powerful tools to advance RNAi analysis

BLOCK-iT Products: Powerful tools to advance RNAi analysis BLOCK-iT RNAi Products BLOCK-iT Products: Powerful tools to advance RNAi analysis With BLOCK-iT RNAi Technology you can: Get powerful blocking and easily generate functional RNAi data Effectively prepare

More information

Genetic Engineering and Biotechnology

Genetic Engineering and Biotechnology 1 So, what is biotechnology?? The use of living organisms to carry out defined chemical processes for industrial or commercial application. The office of Technology Assessment of the U.S. Congress defines

More information

RNA: Transcription and Processing

RNA: Transcription and Processing 8 RNA: Transcription and Processing WORKING WITH THE FIGURES 1. In Figure 8-3, why are the arrows for genes 1 and 2 pointing in opposite directions? The arrows for genes 1 and 2 indicate the direction

More information

RNA Interference. - Localization - Labeling - Expression Profiling - In Vivo Delivery - Transfection

RNA Interference. - Localization - Labeling - Expression Profiling - In Vivo Delivery - Transfection RNA Interference - Localization - Labeling - Expression Profiling - In Vivo Delivery - Transfection Rna interference x RNAi Pathways RNA interference x Tools and Multiplexing RNAi Tools pri-mirna Application

More information

Mouse ES Cell Nucleofector Kit

Mouse ES Cell Nucleofector Kit page 1 of 7 Mouse ES Cell Nucleofector Kit for Mouse Embryonic Stem Cells Cell type Origin Cells derived from mouse blastocysts. Morphology Round cells growing in clumps. Important remarks! 1. This protocol

More information

Gene Silencing Oligos (GSOs) Third Generation Antisense

Gene Silencing Oligos (GSOs) Third Generation Antisense Gene Silencing Oligos (GSOs) Third Generation Antisense Walter R. Strapps, Ph.D. Executive Director, RNA Therapeutics Idera Pharmaceuticals Cambridge, MA NASDAQ: IDRA www.iderapharma.com Idera is a leader

More information

Chapter 7. Physical interaction between Jamb and Jamc is necessary for myoblast fusion. Summary

Chapter 7. Physical interaction between Jamb and Jamc is necessary for myoblast fusion. Summary Chapter 7 Physical interaction between Jamb and Jamc is necessary for myoblast fusion Summary In this chapter I describe a series of transplant experiments designed to further elucidate the function and

More information