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September 10, 2025CellsOpen Access

Rapid and Robust Generation of Homozygous Fluorescent Reporter Knock-In Cell Pools by CRISPR-Cas9

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Authors

JYJicheng YangFGFusheng GuoHCHui San Chin

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Overview

This research demonstrates a novel method to generate stable fluorescent reporter cell pools, indicating improved knock-in efficiency and reduced random integration.

Key Points

  • The study achieved high knock-in efficiency while minimizing random integration through optimized donor DNA design.
  • Using fluorescence-assisted cell sorting, specific homozygous fluorescent clones were successfully isolated and expanded.
  • Methods using integrase-deficient lentivirus vector showed significant advantages over traditional electroporation techniques.
  • The generated reporter cell lines can be utilized for unbiased CRISPR screenings to discover key transcriptional regulators.

Cite This Study

Yang et al. (2025) studied this question.

synapsesocial.com/papers/68c1ad6a54b1d3bfb60e5dd5https://doi.org/10.3390/cells14151165
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