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Generation of a dual edited human induced pluripotent stem cell Myl7-GFP reporter line with inducible CRISPRi/dCas9.
Metzl-Raz, Eyal; Bharucha, Nike; Arthur Ataam, Jennifer; Gavidia, Alexandra A; Greenleaf, William J; Karakikes, Ioannis.
Afiliación
  • Metzl-Raz E; Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
  • Bharucha N; Department Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford, CA, USA.
  • Arthur Ataam J; Department Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford, CA, USA.
  • Gavidia AA; Department Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford, CA, USA.
  • Greenleaf WJ; Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
  • Karakikes I; Department Cardiothoracic Surgery and Stanford Cardiovascular Institute, Stanford, CA, USA.
Stem Cell Res ; 61: 102754, 2022 05.
Article en En | MEDLINE | ID: mdl-35325819
Temporal regulation of CRISPRi activity is critical for genetic screens. Here, we present an inducible CRISPRi platform enabling selection of iPSC-derived cardiomyocytes and reversible gene knockdown. We targeted a doxycycline-inducible dCas9-KRAB-mCherry cassette into the AAVS1 locus in an MYL7-mGFP reporter iPSC line. A clone with bi-allelic integration displayed minimally leaky CRISPRi activity and strong expression upon addition of doxycycline in iPSCs, iPSC-derived cardiomyocytes, and multilineage differentiated cells. The CRISPRi activity was validated by targeting the MYOCD gene in iPSC cardiomyocytes. In summary, we developed a robust inducible CRISPRi platform to interrogate gene function in human iPSC-derived cardiomyocytes and other cells.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes Inducidas Límite: Humans Idioma: En Revista: Stem Cell Res Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes Inducidas Límite: Humans Idioma: En Revista: Stem Cell Res Año: 2022 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido