A conditional counterselectable Piga knockout in mouse embryonic stem cells for advanced genome writing applications.

ABSTRACT: Overwriting counterselectable markers is an efficient strategy for removing wild-type DNA or replacing it with payload DNA of interest. Currently, one bottleneck of efficient genome engineering in mammals is the shortage of counterselectable (negative selection) markers that work robustly without affecting organismal developmental potential. Here, we report a conditional Piga knockout strategy that enables efficient proaerolysin-based counterselection in mouse embryonic stem cells. The conditional Piga knockout cells show similar proaerolysin resistance as full (non-conditional) Piga deletion cells, which enables the use of a PIGA transgene as a counterselectable marker for genome engineering purposes. Native Piga function is readily restored in condition

SUBMITTER: Zhang W 

PROVIDER: S-EPMC9184564 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature