Fast Fission Yeast Genome Editing by CRISPR/Cas9 Using Gap Repair and Fluoride Selection.

Autor:	Ren Y; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.; Nanobiology Institute, Yale University, West Haven, CT, USA., Fernandez R; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.; Nanobiology Institute, Yale University, West Haven, CT, USA.; Bio-X Diagnostics/Adiagène, Ploufragan, France., Saito T; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.; Nanobiology Institute, Yale University, West Haven, CT, USA.; Graduate School of Biomedical Engineering, Tohoku University, Miyagi, Japan., Fujita B; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.; Nanobiology Institute, Yale University, West Haven, CT, USA.; Department of Physiology and Biophysics, Virginia Commonwealth University, Richmond, VA, USA., Mousavi I; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.; Nanobiology Institute, Yale University, West Haven, CT, USA., Berro J; Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA. julien.berro@yale.edu.; Nanobiology Institute, Yale University, West Haven, CT, USA. julien.berro@yale.edu.; Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA. julien.berro@yale.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2025; Vol. 2862, pp. 141-154.
DOI:	10.1007/978-1-0716-4168-2_10
Abstrakt:	We present a protocol to perform CRISPR/Cas9-mediated genome editing in the fission yeast Schizosaccharomyces pombe that does not require cloning and uses the fluoride exporter channel Fex1 as the selection marker. Transformation is typically carried out on the same day of PCR primer arrival and successfully edited strains are selected 5 days after transformation. We expect the adoption of this protocol to further accelerate the throughput of genome editing in S. pombe. (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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