Reference: Gopalakrishnan R, et al. (2025) A comparative review of vector insertion techniques in Saccharomyces cerevisiae. J Microbiol Methods 241:107378

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Abstract


Saccharomyces cerevisiae, a model organism in genetics and molecular biology has been extensively engineered using various vector insertion techniques. This review compares and contrasts three prominent techniques: In vivo homologous recombination (HR), Cre-lox recombination and CRISPR/Cas9. In vivo HR leverages the organism's innate DNA repair machinery for easy vector integration and targeted genome modifications. Cre-lox recombination offers high specificity and efficiency at loxP sites, making it ideal for targeted gene excision or integration. CRISPR/Cas9 has revolutionized genome engineering with its precision and ability to target multiple loci simultaneously. Each technique has its strengths and limitations, including site dependency, off-target effects, and strain-specific variability. This review provides a comprehensive overview of these vector insertion techniques, highlighting their applications, advantages, and limitations in S. cerevisiae genome engineering and synthetic biology.

Reference Type
Journal Article | Review
Authors
Gopalakrishnan R, Kannan K, Gunasekaran R, Ramachandran P, Ganapathy D, Pitchiah S
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