TY - JOUR T1 - Computationally defined and <em>in vitro</em> validated putative genomic safe harbour loci for transgene expression in human cells JF - bioRxiv DO - 10.1101/2021.12.07.471422 SP - 2021.12.07.471422 AU - Matias I. Autio AU - Efthymios Motakis AU - Arnaud Perrin AU - Talal Bin Amin AU - Zenia Tiang AU - Dang Vinh Do AU - Jiaxu Wang AU - Joanna Tan AU - Wei Xuan Tan AU - Adrian Kee Keong Teo AU - Roger S.-Y. Foo Y1 - 2022/01/01 UR - http://biorxiv.org/content/early/2022/01/25/2021.12.07.471422.abstract N2 - Selection of the target site is an inherent question for any project aiming for directed transgene integration. Genomic safe harbour (GSH) loci have been proposed as safe sites in the human genome for transgene integration. Although several sites have been characterised for transgene integration in the literature, most of these do not meet criteria set out for a GSH and the limited set that do have not been characterised extensively. Here, we conducted a computational analysis using publicly available data to identify 25 unique putative GSH loci that reside in active chromosomal compartments. We validated stable transgene expression and minimal disruption of the native transcriptome in three GSH sites in vitro using human embryonic stem cells (hESCs) and their differentiated progeny. Furthermore, for easy targeted transgene expression, we have engineered constitutive landing pad expression constructs into the three validated GSH in hESCs.Competing Interest StatementThe authors have declared no competing interest. ER -