Silencing via lentiviral vectors
Friday, August 1, 2008 at 1:28 pm CDT by David Crotty permalink
August’s issue of CSH Protocols is now available, and one of the featured protocols this month comes from Inder Verma’s lab, and covers the Design and Cloning of an shRNA into a Lentiviral Vector. Combining the specificity of small interfering RNA (siRNA) silencing with the versatility of lentiviral vectors gives researchers a powerful tool for the investigation of gene function both in vivo and in vitro. There’s also an alternative method available. In the featured method, one undesirable consequence of this procedure is that the siRNA target sequence is also present in the mRNA expressing the marker gene, resulting in somewhat lower expression of the marker. In the alternative method, the position of the silencing cassette is upstream of the marker expression cassette, thus avoiding down-regulation of the marker. But, because the silencing cassette is not in the 3′ LTR, only one copy of the silencing cassette is delivered per viral particle (as opposed to two copies in the featured method).
All of our monthly featured articles are freely available to subscribers and non-subscribers alike.
Posted in Cell Biology, DNA Delivery/Gene Transfer, Developmental Biology, General, Genetics, Molecular Biology, RNA Interference (RNAi)/siRNA |
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Sunday, August 3, 2008 at 10:24 am CDT
I suppose it is a lot more work to make up a pdf version? I like to read as if a journal page, i.e., with figures in the body of the paper instead of windows.
Thanks. Jim
Sunday, August 3, 2008 at 7:54 pm CDT
We can’t do pdf’s because the site is set up to work using a common database of recipes and cautions. Each paper links out to that info, which is vital for performing the protocol and is frequently updated (at least this is the case for the cautions).
I would suggest taking a look at the printer-friendly version of any article though, as it gives you everything in one spot, at least a little closer to what you’re looking for. The printer-friendly version link can be found in the information panel at the top right of every article. For the lentivirus one, the link is:
http://www.cshprotocols.org/cgi/content/full/2008/9/pdb.prot5009?print=true