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, the strengths of different classes of site-specific recombinases were merged and combined 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). The authors demonstrated the versatility of STRAIGHT-IN (serine and tyrosine recombinase-assisted integration of genes for high-throughput investigation) by (1) inserting various combinations of fluorescent reporters into hiPSCs to assess the excitation-contraction coupling cascade in derivative cardiomyocytes and (2) simultaneously targeting multiple variants associated with inherited cardiac arrhythmic disorders into a pool of hiPSCs. STRAIGHT-IN offers a precise approach to generate genetically matched panels of hiPSC lines efficiently and cost effectively.
STRAIGHT-IN enables high-throughput targeting of large DNA payloads in human pluripotent stem cells
The IE from MS no longer understands current scripting languages, the latest main version (version 11) is from 2013 and has not been further developed since 2015.
Our recommendation: Use only the latest versions of modern browsers, for example Google Chrome, Mozilla Firefox or Microsofrt Edge, because only this guarantees you sufficient protection against infections and the correct display of websites!