What question did this study set out to answer?

To develop and validate an efficient CRISPR/Cas9-based method for generating targeted large-fragment genomic deletions in human pluripotent stem cells.

October 22, 2025Stem Cell ResearchOpen Access

Targeted large-fragment genomic deletion in human pluripotent stem cells (hPSCs) via CRISPR/Cas9

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Authors

SWSai WeiNanyang Technological University RMRuiting MaFirst Hospital of Lanzhou University TZTing ZhangKunming University of Science and Technology

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Implication

Laboratory protocol demonstrates targeted large-fragment genomic deletion in human pluripotent stem cells, highlighting an efficient approach for functional genomics.

Key Points

To develop and validate an efficient CRISPR/Cas9-based method for generating targeted large-fragment genomic deletions in human pluripotent stem cells.
Designed and synthesized two sgRNAs flanking the targeted genomic region.
Co-delivered sgRNAs with Cas9 protein into single-cell human pluripotent stem cells via electroporation.
Identified deletions by junction PCR, expanded clonal lines, and evaluated stem cell identity, karyotype integrity, and tri-lineage differentiation capacity.
Successfully generated and verified targeted large-fragment genomic deletions in human pluripotent stem cells using a dual-sgRNA CRISPR/Cas9 strategy.
Confirmed that single-cell expanded clones maintained pluripotency markers, intact karyotypes, and full tri-lineage differentiation capacity after editing.

Cite This Study

Wei et al. (2025) studied this question.

synapsesocial.com/papers/6ab384f5acdc7187b41d59e6 https://doi.org/10.1016/j.scr.2025.103859

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Implication

Laboratory protocol demonstrates targeted large-fragment genomic deletion in human pluripotent stem cells, highlighting an efficient approach for functional genomics.

Targeted large-fragment genomic deletion in human pluripotent stem cells (hPSCs) via CRISPR/Cas9

Discussion

Member takes

Implication

Key Points

Cite This Study

Targeted large-fragment genomic deletion in human pluripotent stem cells (hPSCs) via CRISPR/Cas9

Discussion

Member takes

Implication

Key Points

Cite This Study

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