One-step generation of conditional and reversible gene knockouts

作者：Amanda Andersson-Rolf, R. Mustata, A. Merenda, Jihoon Kim, Sajith Perera, Tiago Grego, Katie Andrews, Katie Tremble, José C. R. Silva, Juergen Fink, W. Skarnes, B. Koo · 发表于：Nature Methods · 年份：2017 · DOI：10.1038/nmeth.4156 · 被引用次数：78 · 研究领域：Biology、Medicine

Loss-of-function studies are key for investigating gene function, and 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 biallelic 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.