I"m just wondering if anybody has had any experience with knocking down lncRNAs. I have two lncRNAs of interest that I have knocked down using siRNA, however, I do not see a large knockdown effect. Has anybody else experienced this?
I have just knockout out one using CRISPR/Cas9, and going on knocking, but the mice are still hetero，Be in tough!
I used lenti to express shRNA targeting a lncRNA from the pLKO.1 vector in several mammalian cell lines and found nice knock down.
You can try LNA gapmers. But the knockdown efficiency really depends on the cellular localisation of your lncRNA!.
Hi,
I have a similar problem. When i try to knock down, i see the correct phenotype, the downstream molecule is changed. It seems that knocking down works. However i cannot show how much i knock down of my interested lncrnas. They seem either nit changed or upregulated. I tried many primers. Have you solved the problem? How? Thanks
Regards
You can try complementary LNAs, email me if you need more details.
BW,
a
You could also try siPOOLs. Here's a recent siTOOL's blogpost for a summary of disruption methods - RNAi (with siPOOLs), antisense oligos and CRISPR http://blog.sitoolsbiotech.com/2018/02/disrupting-lncrna-function/
Thanks, I am aware of this method, indeed we discuss it in our recent preprint.
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