In vivo comparative study of RNAi methodologies by in ovo electroporation in the chick embryo

Developmental Dynamics : an Official Publication of the American Association of Anatomists
M RaoShanthini Sockanathan

Abstract

The combination of emergent RNA interference (RNAi) technology with in ovo electroporation in the chick embryo has the potential to provide a powerful and rapid means for functional analyses of novel genes in vivo. In this study, we show that electroporation of short 21-bp RNA duplexes (siRNAs) is a quick and simple method for silencing exogenous and endogenous gene expression in vivo. Quantitative comparisons with two other RNAi protocols that use long double-stranded RNA duplexes and endonuclease-digested duplexes (esiRNAs) demonstrate that siRNAs are significantly more effective at reducing gene expression. Furthermore, we also find that much higher amounts of siRNA are required for silencing of endogenous gene expression relative to plasmid-borne reporter constructs. In short, these results demonstrate that siRNAs are the most effective type of double-stranded RNA duplex for silencing gene expression and suggest that there might be important differences between silencing endogenous and exogenous genes. Finally, we review the parameters for each of these RNA-based methods of RNAi and the controls required to analyze RNAi data in the context of the developing vertebrate embryo.

References

Apr 26, 2001·Developmental Biology·M SwartzC E Krull
May 1, 2001·Methods : a Companion to Methods in Enzymology·H Nakamura, J Funahashi
Mar 9, 2002·Developmental Biology·Janet Heasman
Apr 5, 2002·Advances in Genetics·Alla Grishok, Craig C Mello
Apr 18, 2002·Proceedings of the National Academy of Sciences of the United States of America·Guangchao SuiYang Shi
Jun 11, 2002·Methods : a Companion to Methods in Enzymology·Sayda M ElbashirThomas Tuschl
Oct 3, 2002·Nature Reviews. Genetics·Michael T McManus, Phillip A Sharp
Oct 23, 2002·Proceedings of the National Academy of Sciences of the United States of America·Federico CalegariFrank Buchholz
Dec 24, 2002·Nature Biotechnology·Vladimir PekarikEsther T Stoeckli
Mar 6, 2003·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Robert KosCarol A Erickson
May 20, 2003·Nature Biotechnology·Aimee L JacksonPeter S Linsley
Aug 28, 2003·Nature Cell Biology·Carol A SledzBryan R G Williams
Sep 3, 2003·Development, Growth & Differentiation·Tatsuya Katahira, Harukazu Nakamura
Dec 4, 2003·Nature Biotechnology·Lisa J Scherer, John J Rossi
Feb 6, 2004·Nature Biotechnology·Angela ReynoldsAnastasia Khvorova
Mar 3, 2004·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Catherine E Krull
Jun 2, 2004·Genesis : the Journal of Genetics and Development·Catherine Chesnutt, Lee Niswander

❮ Previous
Next ❯

Citations

May 17, 2005·Nitric Oxide : Biology and Chemistry·Karsten HemmrichVictoria Kolb-Bachofen
Dec 22, 2006·Genes & Development·Maria BergslandJonas Muhr
Jan 18, 2007·Development, Growth & Differentiation·Takahiro YamanishiYuji Yokouchi
Jan 10, 2009·Journal of Dental Research·A H Jheon, R A Schneider
Jan 24, 2006·BMC Developmental Biology·Sanjiv Harpavat, Constance L Cepko
Sep 29, 2007·BMC Developmental Biology·Julien FalkChristine E Holt
Sep 3, 2013·Indian Journal of Virology : an Official Organ of Indian Virological Society·Kathy La Fauce, Leigh Owens
Feb 20, 2013·The Journal of Cell Biology·Kee K KimSachiyo Kawamoto
Sep 21, 2011·Mutation Research·Itzel Sifuentes-RomeroAlejandra García-Gasca
Jul 19, 2005·Domestic Animal Endocrinology·G HenM Friedman-Einat
Sep 30, 2005·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Karine BollérotThierry Jaffredo
Mar 4, 2010·Developmental Biology·Goran PerizShanthini Sockanathan
May 4, 2013·Developmental Cell·Joshua C ChangJane E Johnson
Nov 24, 2016·The Journal of Endocrinology·Joke DelbaereVeerle M Darras
Aug 7, 2007·Cytogenetic and Genome Research·M L V TizardT J Doran
Jun 21, 2005·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Jiankai Luo, Christoph Redies
Dec 8, 2005·Birth Defects Research. Part A, Clinical and Molecular Teratology·Victoria J DrakeSusan M Smith
Mar 12, 2020·Journal of Cardiovascular Development and Disease·Rusty Lansford, Sandra Rugonyi
Apr 20, 2005·Birth Defects Research. Part C, Embryo Today : Reviews·Meenakshi Rao, Shanthini Sockanathan
Oct 1, 2005·Science·Meenakshi Rao, Shanthini Sockanathan
Jan 1, 2008·CSH Protocols·Olivier MautiEsther T Stoeckli

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Mechanisms of Development
Harukazu NakamuraJun-Ichi Funahashi
Proceedings of the National Academy of Sciences of the United States of America
Guangchao SuiYang Shi
Current Opinion in Neurobiology
Amy W Helms, Jane E Johnson
© 2021 Meta ULC. All rights reserved