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Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line

Prime editing uses the Cas9 nickase fused to a reverse transcriptase to copy a DNA sequence into a specific locus from a ‘prime editing’ guide RNA (pegRNA), eliminating the need for double-stranded DNA breaks and donor DNA templates. To facilitate prime editing in human induced pluripotent stem cell...

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Detalles Bibliográficos
Autores principales: Bharucha, Nike, Ataam, Jennifer Arthur, Gavidia, Alexandra A., Karakikes, Ioannis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9126997/
https://www.ncbi.nlm.nih.gov/pubmed/34875545
http://dx.doi.org/10.1016/j.scr.2021.102610
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author Bharucha, Nike
Ataam, Jennifer Arthur
Gavidia, Alexandra A.
Karakikes, Ioannis
author_facet Bharucha, Nike
Ataam, Jennifer Arthur
Gavidia, Alexandra A.
Karakikes, Ioannis
author_sort Bharucha, Nike
collection PubMed
description Prime editing uses the Cas9 nickase fused to a reverse transcriptase to copy a DNA sequence into a specific locus from a ‘prime editing’ guide RNA (pegRNA), eliminating the need for double-stranded DNA breaks and donor DNA templates. To facilitate prime editing in human induced pluripotent stem cells (iPSCs), we integrated a doxycycline-inducible Prime Editor protein (PE2) into the AAVS1 genomic safe harbor locus. Prime editing of iPSCs resulted in precise insertion of three nucleotides in HEK3 locus with high efficiency, demonstrating the utility of this approach. This engineered cell line can be used to edit a single or multiple genomic loci by introducing a target-specific pegRNA for precise and effective genome editing to facilitate disease modeling and functional genetics studies.
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spelling pubmed-91269972023-05-24 Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line Bharucha, Nike Ataam, Jennifer Arthur Gavidia, Alexandra A. Karakikes, Ioannis Stem Cell Res Article Prime editing uses the Cas9 nickase fused to a reverse transcriptase to copy a DNA sequence into a specific locus from a ‘prime editing’ guide RNA (pegRNA), eliminating the need for double-stranded DNA breaks and donor DNA templates. To facilitate prime editing in human induced pluripotent stem cells (iPSCs), we integrated a doxycycline-inducible Prime Editor protein (PE2) into the AAVS1 genomic safe harbor locus. Prime editing of iPSCs resulted in precise insertion of three nucleotides in HEK3 locus with high efficiency, demonstrating the utility of this approach. This engineered cell line can be used to edit a single or multiple genomic loci by introducing a target-specific pegRNA for precise and effective genome editing to facilitate disease modeling and functional genetics studies. 2021-11-24 /pmc/articles/PMC9126997/ /pubmed/34875545 http://dx.doi.org/10.1016/j.scr.2021.102610 Text en https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Bharucha, Nike
Ataam, Jennifer Arthur
Gavidia, Alexandra A.
Karakikes, Ioannis
Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title_full Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title_fullStr Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title_full_unstemmed Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title_short Generation of AAVS1 integrated doxycycline-inducible CRISPR-Prime Editor human induced pluripotent stem cell line
title_sort generation of aavs1 integrated doxycycline-inducible crispr-prime editor human induced pluripotent stem cell line
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9126997/
https://www.ncbi.nlm.nih.gov/pubmed/34875545
http://dx.doi.org/10.1016/j.scr.2021.102610
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