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Eng"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Targeting DNA payloads into human induced pluripotent stem cells (hiPSCs) typically requires multiple inefficient steps, slowing the testing of gene circuits and cell-fate programmes. Here we show that STRAIGHT-IN Dual enables simultaneous, allele-specific, single-copy integration of two DNA constructs efficiently within 1 week. STRAIGHT-IN Dual leverages the STRAIGHT-IN platform for near-scarless payload integration, facilitating the recycling of components for further modifications. Using STRAIGHT-IN Dual, we investigate how promoter choice and gene syntax influence transgene silencing and how these design features affect reporter expression and forward programming of hiPSCs into neurons, motor neurons and endothelial cells. 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STRAIGHT-IN Dual generates homogeneous engineered hiPSC populations, accelerating synthetic biology design\u2013build\u2013test cycles in stem cells and enabling controlled comparisons of circuit performances.<\/jats:p>","DOI":"10.1038\/s41551-026-01677-9","type":"journal-article","created":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T09:02:28Z","timestamp":1777539748000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["STRAIGHT-IN Dual: a platform for dual single-copy integrations of DNA payloads and gene circuits into human induced pluripotent stem cells"],"prefix":"10.1038","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5608-220X","authenticated-orcid":false,"given":"Albert","family":"Blanch-Asensio","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5033-2068","authenticated-orcid":false,"given":"Deon S.","family":"Ploessl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6280-8385","authenticated-orcid":false,"given":"Benjamin B.","family":"Johnson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Cascione","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3217-5676","authenticated-orcid":false,"given":"Myrthe R. 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