Computationally defined and in vitro validated putative genomic safe harbour loci for transgene expression in human cells.

Autor:	Autio MI; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore.; Laboratory of Systems Biology and Data Analytics, Genome Institute of Singapore, Singapore, Singapore., Motakis E; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore., Perrin A; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore., Bin Amin T; Laboratory of Systems Biology and Data Analytics, Genome Institute of Singapore, Singapore, Singapore., Tiang Z; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore., Do DV; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore., Wang J; Laboratory of RNA Genomics and Structure, Genome Institute of Singapore, Singapore, Singapore., Tan J; Center for Genome Diagnostics, Genome Institute of Singapore, Singapore, Singapore., Ding SSL; Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, Singapore, Singapore., Tan WX; Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, Singapore, Singapore.; Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore., Lee CJM; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore., Teo AKK; Stem Cells and Diabetes Laboratory, Institute of Molecular and Cell Biology, Singapore, Singapore.; Department of Medicine, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.; Precision Medicine Translational Research Programme, Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore., Foo RSY; Laboratory of Molecular Epigenomics and Chromatin Organization, Genome Institute of Singapore, Singapore, Singapore.; Cardiovascular Disease Translational Research Programme, Yong Loo Lin School of Medicine, Singapore, Singapore.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2024 Jan 02; Vol. 13. Date of Electronic Publication: 2024 Jan 02.
DOI:	10.7554/eLife.79592
Abstrakt:	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 Interests: MA, EM, AP, RF Patent application PCT/SG2022/050888, TB, ZT, DD, JW, JT, SD, WT, CL, AT No competing interests declared (© 2024, Autio et al.)
Databáze:	MEDLINE
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