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In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
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Efficient gene editing through an intronic selection marker in cells.

Shang Wang1,2, Yuqing Li2,3, Li Zhong4,5

  • 1State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Department of Experimental Research, Sun Yat-Sen University Cancer Center, Guangzhou, 510060, China.

Cellular and Molecular Life Sciences : CMLS
|January 31, 2022
PubMed
Summary
This summary is machine-generated.

Researchers developed Gene Editing through an Intronic Selection marker (GEIS), a new HDR gene editing strategy. GEIS improves DNA variation editing efficiency and accuracy in eukaryotic cells, offering a promising tool for genetic research and therapy.

Keywords:
Conversion tractFACSGene editingHDRIntron reporter

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Gene editing enables genome modification in eukaryotes for research and therapy.
  • Current limitations include low homology-directed DNA repair (HDR) efficiency and inability of base editors (BEs) to perform indel editing.

Purpose of the Study:

  • To improve HDR gene editing technology for efficient and accurate DNA variation introduction.
  • To develop a novel strategy that overcomes limitations of existing gene editing methods.

Main Methods:

  • Development of a new HDR strategy named Gene Editing through an Intronic Selection marker (GEIS).
  • Utilized an independent selection marker within an intron for HDR-mediated DNA variation.
  • Employed fluorescence-activated cell sorting (FACS) for selection and validation.

Main Results:

  • GEIS achieved a DsRed positive rate of 87.5% after two FACS selections.
  • The strategy effectively avoids nonhomologous end joining (NHEJ)-mediated false positives.
  • GEIS demonstrated potential to edit approximately 97% of gene editing targets in human and mouse cells without affecting endogenous gene expression.

Conclusions:

  • GEIS represents a significant advancement in HDR gene editing technology.
  • The strategy offers high efficiency and accuracy for introducing DNA variations.
  • GEIS shows potential for editing multiple DNA variations and broad applicability in human and mouse cells.