I am just wondering if this is possible and all the publications I have come across appear to knockdown ATRX using siRNA or shRNA but they never report how long these knockdowns last. Any input would be great.
Hi Sophie,
If you are using cell lines, you should be able to make a stable line by transfecting your shRNA plasmid and then selecting stable clones, like you would make a stable line overexpressing a given protein. You can then assess the efficiency of the knockdown by western blot. This has been discussed in a previous RG thread that I would recommend https://www.researchgate.net/post/Transient_and_stable_transfections_by_shRNA_Is_it_just_a_matter_of_time
Hope this help. Good luck!
I guess, siRNA effect does not last long as it seems to wear off after 96 hours. shRNA mediated gene silencing seems to be a good option than siRNA. You can transfect cells with shRNA and select clone. As Norbert said earlier, you need to check KD level at protein level. If you are looking for a complete gene knockout, then CRISPR/CAS9 system will be very useful.
I was able to get a successful stable knockdowns for a same gene in both human and mouse epithelial cell lines using Sigma lentiviral particles. They sell these either in plasmid form or as transduction particles which are ready to use. Please see the link below for your gene. Good luck!
http://www.sigmaaldrich.com/catalog/genes/ATRX?lang=en®ion=US#shRNA%20Products~human
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