Efficient and flexible tagging of endogenous genes by homology-independent intron targeting.

ABSTRACT: Genome editing tools have simplified the generation of knock-in gene fusions, yet the prevalent use of gene-specific homology-directed repair (HDR) templates still hinders scalability. Consequently, realization of large-scale gene tagging requires further development of approaches to generate knock-in protein fusions via generic donors that do not require locus-specific homology sequences. Here, we combine intron-based protein trapping with homology-independent repair-based integration of a generic donor and demonstrate precise, scalable, and efficient gene tagging. Because editing is performed in introns using a synthetic exon, this approach tolerates mutations in the unedited allele, indels at the integration site, and the addition of resistance genes that do not disrupt the target gene

SUBMITTER: Serebrenik YV 

PROVIDER: S-EPMC6673721 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature