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Establishment of a second generation homozygous CRISPRa human induced pluripotent stem cell (hiPSC) line for enhanced levels of endogenous gene activation.

Research paper by Eric E Schoger, Wolfram-Hubertus WH Zimmermann, Lukas L Cyganek, Laura Cecilia LC Zelarayán

Indexed on: 05 Sep '21Published on: 05 Sep '21Published in: Stem Cell Research



Abstract

CRISPR/Cas9 technology based on nuclease inactive dCas9 and fused to the heterotrimeric VPR transcriptional activator is a powerful tool to enhance endogenous transcription by targeting defined genomic loci. We generated homozygous human induced pluripotent stem cell (hiPSC) lines carrying dCas9 fused to VPR along with a WPRE element at the AAVS1 locus (CRISPRa2). We demonstrated pluripotency, genomic integrity and differentiation potential into all three germ layers. CRISPRa2 cells showed increased transgene expression and higher transcriptional induction in hiPSC-derived cardiomyocytes compared to a previously described CRISPRa line. Both lines allow studying endogenous transcriptional modulation with lower and higher transcript abundance. Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.