Designing CRISPR-Cas9 Constructs to Target the SIFSR Gene in Tomato Varieties Pusa Ruby and UC82 for Extended Shelf Life

Abstract

This research focuses on the strategic design and implementation of CRISPR-Cas9 constructs to target the Solanum lycopersicum Shelf-Life Food Regulator (SlFSR) gene in two tomato varieties, Pusa Ruby and UC82, to extend post-harvest shelf life. Precise gene editing was done by meticulously selecting target sites within the SIFSR gene and optimizing guide RNA sequences. The CRISPR-Cas9 cassettes were PCR-amplified, cloned into vectors, and ligated to generate the final constructs. Subsequently, these constructs were introduced into both tomato varieties using appropriate delivery methods and transformation protocols. Our findings underscore the importance of precision engineering techniques such as cloning, PCR, and ligation in leveraging genetic editing for agricultural purposes. This research contributes insights into refining CRISPR-Cas9 technology for targeted genetic modification, particularly in enhancing the shelf life of fresh produce, thereby promoting sustainable food practices.