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CRISPR interference is a method to conditionally inhibit transcription of an arbitrary target gene. This is useful to study the functions of essential genes, which are required for cellular viability. Although many conditional gene perturbation techniques are available for Schizosaccharomyces pombe, CRISPRi facilitates construction of a large number of knockdown strains because of its systematic, simple procedure. Here, we describe a method to construct and characterize knockdown strains using dCas9-mediated CRISPRi in S. pombe, including a variation of CRISPRi induction technique in a 96-well format for high-throughput studies.

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Proliferation Analyses of Conditional Knockdown Strains Using CRISPR Interference in Fission Yeast

  • 实验方案

  • 发表日期：

    2025年

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  • Ken Ishikawa,
  • Shigeaki Saitoh

摘要

CRISPR interference is a method to conditionally inhibit transcription of an arbitrary target gene. This is useful to study the functions of essential genes, which are required for cellular viability. Although many conditional gene perturbation techniques are available for Schizosaccharomyces pombe, CRISPRi facilitates construction of a large number of knockdown strains because of its systematic, simple procedure. Here, we describe a method to construct and characterize knockdown strains using dCas9-mediated CRISPRi in S. pombe, including a variation of CRISPRi induction technique in a 96-well format for high-throughput studies.