A high efficiency precision genome editing method with CRISPR in iPSCs

Avinash Singh; G. Dalton Smedley; Jamee Grace Rose; Kristina Fredriksen; Ying Zhang; Ling Li; Shauna H. Yuan

doi:10.1038/s41598-024-60766-4

A high efficiency precision genome editing method with CRISPR in iPSCs

Avinash Singh, G. Dalton Smedley, Jamee Grace Rose, Kristina Fredriksen, Ying Zhang, Ling Li, Shauna H. Yuan

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

The use of genetic engineering to generate point mutations in induced pluripotent stem cells (iPSCs) is essential for studying a specific genetic effect in an isogenic background. We demonstrate that a combination of p53 inhibition and pro-survival small molecules achieves a homologous recombination rate higher than 90% using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in human iPSCs. Our protocol reduces the effort and time required to create isogenic lines.

Original language	English (US)
Article number	9933
Journal	Scientific reports
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41598-024-60766-4
State	Published - Dec 2024

Bibliographical note

Publisher Copyright:
© The Author(s) 2024.

Keywords

CRISPR
Gene editing
High efficiency
Single nucleotide polymorphism
iPSC

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

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10.1038/s41598-024-60766-4

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AU - Singh, Avinash

AU - Smedley, G. Dalton

AU - Rose, Jamee Grace

AU - Fredriksen, Kristina

AU - Zhang, Ying

AU - Li, Ling

AU - Yuan, Shauna H.

N1 - Publisher Copyright: © The Author(s) 2024.

PY - 2024/12

Y1 - 2024/12

N2 - The use of genetic engineering to generate point mutations in induced pluripotent stem cells (iPSCs) is essential for studying a specific genetic effect in an isogenic background. We demonstrate that a combination of p53 inhibition and pro-survival small molecules achieves a homologous recombination rate higher than 90% using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in human iPSCs. Our protocol reduces the effort and time required to create isogenic lines.

AB - The use of genetic engineering to generate point mutations in induced pluripotent stem cells (iPSCs) is essential for studying a specific genetic effect in an isogenic background. We demonstrate that a combination of p53 inhibition and pro-survival small molecules achieves a homologous recombination rate higher than 90% using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) in human iPSCs. Our protocol reduces the effort and time required to create isogenic lines.

KW - CRISPR

KW - Gene editing

KW - High efficiency

KW - Single nucleotide polymorphism

KW - iPSC

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DO - 10.1038/s41598-024-60766-4

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SN - 2045-2322

VL - 14

JO - Scientific reports

JF - Scientific reports

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ER -

A high efficiency precision genome editing method with CRISPR in iPSCs

Abstract

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Keywords

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