One-step generation of conditional and reversible gene knockouts.

Abstract

CRISPR technology has made genome editing widely accessible in model organisms and cells. However, conditional gene inactivation in diploid cells is still difficult to achieve. Here, we present CRISPR-FLIP, a strategy that provides an efficient, rapid, and scalable method for bi-allelic conditional gene knockouts in diploid or aneuploid cells such as pluripotent stem cells, 3D organoids and cell lines by co-delivery of CRISPR/Cas9 and a universal conditional intronic cassette.