DNA barcoding reveals that injected transgenes are predominantly processed by homologous recombination in mouse zygote.

ABSTRACT: Mechanisms that ensure repair of double-strand DNA breaks (DSBs) are instrumental in the integration of foreign DNA into the genome of transgenic organisms. After pronuclear microinjection, exogenous DNA is usually found as a concatemer comprising multiple co-integrated transgene copies. Here, we investigated the contribution of various DSB repair pathways to the concatemer formation. We injected mouse zygotes with a pool of linear DNA molecules carrying unique barcodes at both ends and obtained 10 transgenic embryos with 1-300 transgene copies. Sequencing the barcodes allowed us to assign relative positions to the copies in concatemers and detect recombination events that occurred during integration. Cumulative analysis of approximately 1,000 integrated copies reveals that over 80% of the

SUBMITTER: Smirnov A 

PROVIDER: S-EPMC7145541 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature