Scientific article
English

A toolbox of Stable Integration Vectors (SIV) in the fission yeast Schizosaccharomyces pombe

Published inJournal of cell science, vol. 133, jcs.240754
Publication date2019-01-01
First online date2019-01-01
Abstract

Schizosaccharomyces pombe is a widely used model organism to study many aspects of eukaryotic cell physiology. Its popularity as an experimental system partially stems from the ease of genetic manipulations, where the innate homology-targeted repair is exploited to precisely edit the genome. While vectors to incorporate exogenous sequences into the chromosomes are available, most are poorly characterized. Here we show that commonly used fission yeast vectors, which upon integration produce repetitive genomic regions, yield unstable genomic loci. We overcome this problem by designing a new series of Stable Integration Vectors (SIV) that target four different prototrophy genes. SIV produce non-repetitive, stable genomic loci and integrate predominantly as single copy. Additionally, we develop a set of complementary auxotrophic alleles that preclude false-positive integration events. We expand the vector series to include antibiotic resistance markers, promoters, fluorescent tags and terminators, and build a highly modular toolbox to introduce heterologous sequences. Finally, as proof of concept, we generate a large set of ready-to-use, fluorescent probes to mark organelles and cellular processes with a wide range of applications in fission yeast research.

Keywords
  • Fission yeast
  • Genome stability
  • Integrative vector
  • Molecular genetics
  • Schizosaccharomyces pombe
Affiliation entities Not a UNIGE publication
Citation (ISO format)
VJEŠTICA, Aleksandar et al. A toolbox of Stable Integration Vectors (SIV) in the fission yeast Schizosaccharomyces pombe. In: Journal of cell science, 2019, vol. 133, p. jcs.240754. doi: 10.1242/jcs.240754
Main files (1)
Article (Published version)
accessLevelRestricted
Identifiers
Journal ISSN0021-9533
59views
0downloads

Technical informations

Creation30/01/2024 17:30:04
First validation31/01/2024 15:54:58
Update31/01/2024 15:54:58
Status update31/01/2024 15:54:58
Last indexation01/11/2024 07:22:45
All rights reserved by Archive ouverte UNIGE and the University of GenevaunigeBlack