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

Ken Ishikawa; Shigeaki Saitoh

doi:10.1007/978-1-0716-4168-2_12

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

Methods Mol Biol. 2025:2862:171-186. doi: 10.1007/978-1-0716-4168-2_12.

Authors

Ken Ishikawa¹, Shigeaki Saitoh²

Affiliations

¹ Department of Cell Biology, Institute of Life Science, Kurume University, Kurume, Japan. [email protected].
² Department of Cell Biology, Institute of Life Science, Kurume University, Kurume, Japan.

PMID: 39527200
DOI: 10.1007/978-1-0716-4168-2_12

Abstract

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.

Keywords: CRISPRi; Essential gene; Gene perturbation; Knockdown; Transcription; dCas9.

MeSH terms

CRISPR-Cas Systems*
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Gene Knockdown Techniques* / methods
RNA, Guide, CRISPR-Cas Systems / genetics
Schizosaccharomyces* / genetics

Substances

RNA, Guide, CRISPR-Cas Systems