Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology

Cells

Said Assou¹  Chloé Bourguignon¹  Chloé Nguyen Van¹  Joffrey Mianné¹  Mathieu Fieldès¹  Amel Nasri¹  John De Vos¹ 

[1] IRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, France;

关键词: hPSC;    CRISPR;    screening;    stem cells;    gene editing;    gene targeting;   

DOI  :  10.3390/cells9051312

来源: DOAJ

【 摘 要 】

Recent advances in genome engineering based on the CRISPR/Cas9 technology have revolutionized our ability to manipulate genomic DNA. Its use in human pluripotent stem cells (hPSCs) has allowed a wide range of mutant cell lines to be obtained at an unprecedented rate. The combination of these two groundbreaking technologies has tremendous potential, from disease modeling to stem cell-based therapies. However, the generation, screening and molecular characterization of these cell lines remain a cumbersome and multi-step endeavor. Here, we propose a pipeline of strategies to efficiently generate, sub-clone, and characterize CRISPR/Cas9-edited hPSC lines in the function of the introduced mutation (indels, point mutations, insertion of large constructs, deletions).

【 授权许可】

Unknown