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Updated: Oct 27, 2025

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System
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CRISPRi/a Screening with Human iPSCs.

Masataka Nishiga1, Lei S Qi2,3,4, Joseph C Wu5,6,7

  • 1Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 24, 2021
PubMed
Summary
This summary is machine-generated.

This study presents a CRISPR screening protocol using human induced pluripotent stem cells (iPSCs). This method enables the identification of essential genes for differentiating iPSCs into cardiomyocytes (CMs).

Keywords:
CRISPR/Cas9CardiomyocytesGenome editingInduced pluripotent stem cells

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Area of Science:

  • Stem cell biology
  • Genetics and genomics
  • Molecular and cellular biology

Background:

  • Identifying causative genes is crucial for biological discovery and drug development.
  • CRISPR/Cas9 screens are powerful for genetic perturbation but challenging in primary human somatic cells.
  • Induced pluripotent stem cells (iPSCs) offer a scalable solution for CRISPR screening due to their expansion and differentiation potential.

Purpose of the Study:

  • To develop a robust protocol for CRISPR screening using human iPSCs.
  • To demonstrate the utility of this approach for identifying essential genes in cell differentiation.
  • To overcome limitations of CRISPR screening in non-cancerous human somatic cells.

Main Methods:

  • Development of a CRISPR screening protocol tailored for human iPSCs.
  • Implementation of a CRISPR interference (CRISPRi) knockdown screen.
  • Application of the screen to identify genes essential for iPSC differentiation into cardiomyocytes (CMs).

Main Results:

  • Successful establishment of a CRISPR screening platform using human iPSCs.
  • Identification of key genes required for efficient iPSC to CM differentiation.
  • Demonstration of the protocol's adaptability for various cell types and phenotypes.

Conclusions:

  • CRISPR screening with human iPSCs is a powerful and versatile tool for gene discovery.
  • This technology facilitates the identification of causative genes and pathways in various biological processes.
  • The developed protocol overcomes previous limitations, enabling broader application of CRISPR screens in regenerative medicine and disease modeling.