2. Academic Validation
  3. Targeted large-fragment genomic deletion in human pluripotent stem cells (hPSCs) via CRISPR/Cas9

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

  • Stem Cell Res. 2025 Oct 21:89:103859. doi: 10.1016/j.scr.2025.103859.
Sai Wei 1,  Ruiting Ma 2,  Ting Zhang 3,  Luan Wen 4
Affiliations

Affiliations

  • 1 Key Laboratory of Tropical Translational Medicine of Ministry of Education & Key Laboratory of Brain Science Research Transformation in Tropical Environment of Hainan Province, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Hainan Medical University, Haikou, Hainan 571199, China; Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Key Laboratory of Reproductive Health Diseases Research and Translation (Hainan Medical University), Ministry of Education, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, Hainan 571101, China.
  • 2 Key Laboratory of Tropical Translational Medicine of Ministry of Education & Key Laboratory of Brain Science Research Transformation in Tropical Environment of Hainan Province, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Hainan Medical University, Haikou, Hainan 571199, China.
  • 3 School of Tropical Medicine, Hainan Medical University, Haikou, Hainan 571199, China. Electronic address: hy0211105@muhn.edu.cn.
  • 4 Key Laboratory of Tropical Translational Medicine of Ministry of Education & Key Laboratory of Brain Science Research Transformation in Tropical Environment of Hainan Province, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Hainan Medical University, Haikou, Hainan 571199, China; Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Key Laboratory of Reproductive Health Diseases Research and Translation (Hainan Medical University), Ministry of Education, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, Hainan 571101, China. Electronic address: luanwen@muhn.edu.cn.
PMID: 41167048 DOI: 10.1016/j.scr.2025.103859
Abstract

Large genomic deletions are increasingly recognized as contributors to human disease. Here, we describe a CRISPR/Cas9-based protocol to induce targeted large-fragment deletions in human pluripotent stem cells (hPSCs). Two sgRNAs flanking the target region were designed, synthesized, and co-delivered with Cas9 protein into single-cell hPSCs via electroporation. Deletions were identified using junction PCR. Successfully edited cell pools were expanded as single-cell colonies and assessed for genomic deletion, stem cell identity, karyotype integrity, and tri-lineage differentiation capacity. This approach provides a robust method for modeling genomic deletions in hPSCs for Disease Research and functional genomics.

Keywords

CRISPR/Cas9; Large-fragment deletion editing; RNP.

Figures
Products
嗨！很高兴为您提供帮助！

尊敬的 MCE 客户您好，
请您选择所在区域，我们将转接对应客服为您服务！

热门区域

A

B

C

F

G

H

J

L

N

Q

S

T

X

Y

Z

热 A B C F G H J L N Q S T X Y Z