title: High-Content Analysis of CRISPR-Cas9 Gene-Edited Human Embryonic Stem Cells.
abstract: CRISPR-Cas9 gene editing of human cells and tissues holds much promise to advance medicine and biology, but standard editing methods require weeks to months of reagent preparation and selection where much or all of the initial edited samples are destroyed during analysis. ArrayEdit, a simple approach utilizing surface-modified multiwell plates containing one-pot transcribed single-guide RNAs, separates thousands of edited cell populations for automated, live, high-content imaging and analysis. The approach lowers the time and cost of gene editing and produces edited human embryonic stem cells at high efficiencies. Edited genes can be expressed in both pluripotent stem cells and differentiated cells. This preclinical platform adds important capabilities to observe editing and selection in situ within complex structures generated by human cells, ultimately enabling optical and other molecular perturbations in the editing workflow that could refine the specificity and versatility of gene editing.
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Jared
authors.initials: J
authors.lastname: Carlson-Stevermer
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Madelyn
authors.initials: M
authors.lastname: Goedland
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Steyer
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Arezoo
authors.initials: A
authors.lastname: Movaghar
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Meng
authors.initials: M
authors.lastname: Lou
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Lucille
authors.initials: L
authors.lastname: Kohlenberg
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Prestil
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Medical History and Bioethics, University of Wisconsin-Madison, Madison, WI 53715, USA. Electronic address: ksaha@wisc.edu.
authors.forename: Krishanu
authors.initials: K
authors.lastname: Saha
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D018390:Gene Targeting / Q000295:instrumentation / Q000379:methods*; D015894:Genome, Human / Q000235:genetics*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors