STRAIGHT-IN enables high-throughput targeting of large DNA payloads in human pluripotent stem cells.

ABSTRACT: Inserting large DNA payloads (>10 kb) into specific genomic sites of mammalian cells remains challenging. Applications ranging from synthetic biology to evaluating the pathogenicity of disease-associated variants for precision medicine initiatives would greatly benefit from tools that facilitate this process. Here, we merge the strengths of different classes of site-specific recombinases and combine these with CRISPR-Cas9-mediated homologous recombination to develop a strategy for stringent site-specific replacement of genomic fragments at least 50 kb in size in human induced pluripotent stem cells (hiPSCs). We demonstrate the versatility of STRAIGHT-IN (serine and tyrosine recombinase-assisted integration of genes for high-throughput investigation) by (1) inserting various combinations of

SUBMITTER: Blanch-Asensio A 

PROVIDER: S-EPMC9606106 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature