Arrayed mutant haploid embryonic stem cell libraries facilitate phenotype-driven genetic screens.

ABSTRACT: Forward genetic screens using mammalian embryonic stem (ES) cells have identified genes required for numerous cellular processes. However, loss-of-function screens are more difficult to conduct in diploid cells because, in most cases, both alleles of a gene must be mutated to exhibit a phenotype. Recently, mammalian haploid ES cell lines were successfully established and applied to several recessive genetic screens. However, all these screens were performed in mixed pools of mutant cells and were mainly based on positive selection. In general, negative screening is not easy to apply to these mixed pools, although quantitative deep sequencing of mutagen insertions can help to identify some 'missing' mutants. Moreover, the interplay between different mutant cells in the mixed pools would int

SUBMITTER: Liu G 

PROVIDER: S-EPMC5727442 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature