RNAi methods updated
Wednesday, January 21, 2009 at 10:01 am CST by David Crotty permalink
Way back in 2003, we published RNAi: A Guide To Gene Silencing, which was one of, if not the first major treatises on the subject. One of the problems with being the first to publish on a fast-moving field is that a book can date quickly. While there’s still much valuable information in RNAi, I’ve been asking authors to update their protocols, which have evolved over the last 5 years or so.
Last month, Esther Stoeckli and colleagues provided an update to her method for Gene Silencing by Injection and Electroporation of dsRNA in Avian Embryos.
This month’s issue brings a tour de force updating and expansion of Petr Svoboda and Paula Stein’s chapter on RNAi in mouse oocytes and early embryos. They’ve written up a general topic introduction on the subject, explanations of how to choose the sequence of dsRNA for RNAi and how to clone and sequence an inverted repeat, and protocols for Cloning a Transgene for Transgenic RNAi in Mouse Oocytes, Preparation of dsRNA for Microinjection, Microinjection of dsRNA into Fully-Grown Mouse Oocytes, Microinjection of dsRNA into Mouse One-Cell Embryos, and Microinjection of Plasmids into Meiotically Incompetent Mouse Oocytes.
Next month will bring an update of Savithramma Dinesh-Kumar’s protocol for using viral vectors for RNAi in plants. More on that in February.
Posted in Cell Biology, DNA Delivery/Gene Transfer, Developmental Biology, General, Laboratory Organisms, Molecular Biology, RNA Interference (RNAi)/siRNA | 2 Comments »
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Thursday, January 22, 2009 at 10:41 am CST
Thanks for providing this summary of updates–and of course for the updates themselves.
Thursday, January 22, 2009 at 10:48 am CST
Hi Stephanie,
Just to give you the complete picture, a few months back, Bruce Paterson’s lab put together updated versions of protocols for fly RNAi from his earlier chapter in Drosophila Protocols:
Drosophila RNA Interference (RNAi) Using a Gal-4 Inducible Transgene Vector
Preparation of Double-Stranded RNA for Drosophila RNA Interference (RNAi)
Collection of Drosophila Embryos for RNA Interference (RNAi)
and
Injection of dsRNA into Drosophila Embryos for RNA Interference (RNAi)