Expanding The Capabilities Of Targeted Integration

Read details on a major advance in genome engineering: the successful knock‑in of a 50 kb, multi‑functional DNA construct into human induced pluripotent stem cells at a single, defined genomic locus. Traditional methods such as CRISPR‑based homology‑directed repair or viral delivery struggle with large payloads, often sacrificing efficiency, specificity, or expression control. Here, a site‑specific recombinase system enabled reliable integration and consistent expression of multiple constitutive and inducible genes within one engineered cell line. Rapid synthesis of ultra‑long DNA constructs further shortened development timelines, enabling assembly of complex payloads in weeks rather than months. Molecular and functional analyses confirmed high integration efficiency, robust constitutive expression, and inducible control of selected genes. Together, these results expand what is possible for advanced applications such as screening libraries, cell and gene therapy development, multi‑protein expression, and humanized disease models requiring large, precisely engineered genomic insertions