I would like to know information about esiRNA form Sigma for the following points: 1. transfection protocol; 2. knockdown efficiency; 3. off target effects; 4. toxicity. Anyone would like to share the experience? Thanks!
Hi Wensheng, both reagents unfortunately still suffer from off-target effects from our experience.
We find defined, high complexity pools of ~30 siRNAs (siPOOLs https://www.sitoolsbiotech.com/sipools.php ) to work best (I admit this may be a little biased because I work for the cpy that makes them!) But in essence, studies have shown that single siRNAs/low complexity pools of 3-4 siRNAs suffer greatly from off-targets which can compromise results.
Although the esiRNAs are highly complex, the problem with them are their ill-defined lengths. This is because they are generated by a non-specific endoribonuclease that generates heterogenous dsRNA fragments. The spin-column purification used fails to remove longer dsRNAs of ~40 nt which can stimulate immune responses. We've tested and published this here: Article SiPools: Highly complex but accurately defined siRNA pools e...
Smartpools contain 4 siRNAs, which are insufficient to effectively dilute off-target effects. And in fact have been shown to make things worse: http://blog.sitoolsbiotech.com/2018/04/low-complexity-pooling-not-prevent-sirna-off-targets/
For more info on siPOOLs, you can refer to this technical brochure https://www.sitoolsbiotech.com/pdf/170926_siPOOLsProductBrochure2017_webPDF.pdf
If you have more questions, don't hesitate to contact me! Gd luck!
Hi Catherine, thank you very much! I remember your presentation. I will contact you.
- Badges
- Science method