I am considering whether to purchase H9 or H9 Cre-LoxP cells. I know that H9 is a very popular cell line but haven't seen many references for the Cre-LoxP-modified version. Does anyone have experience using the modified line? I'm concerned about whether the LoxP cassette may have randomly inserted into a location that would affect the regulation of genes important to my studies.
Thanks!
I don't have experience using the cell line but based on WiCell declaration, With Cre-mediated recombination, the new transgene can replace the master vector at the same integration site, which guarantees the silence-resistant expression of the new transgene as that of GFP. I suggest you for further and detailed information have a conversation with Dr. Su-Chun Zhang at university of Wisconsin.
Good luck,
Hadi
Hi Stacy, I'm now conducting this kind of work. You can insert Loxp cassette into almost any locus with CRISPR/Cas9 and screen for positive clones without random insertion by Southern blot.
Dear Su
Are you sure about it? if you try to deliver your system by plasmid; random insertion is a limitation even for CRISPR/Cas9 which is observed tolerated up to 5 mismatches in crRNA ( Kim et al. 2014). delivering purified Cas9 has higher accuracy,however.
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