TL;DR
This protocol covers both single-gRNA (sgRNA) and dual-gRNA directed knockout systems and describes the procedure for constructing an Auxin-Inducible Degron (AID) knock-in-mediated system to induce the rapid degradation of a target protein in cells, thereby investigating its function.
This chapter outlines a comprehensive methodology for the design, assembly, and functional assessment of CRISPR/dCas9 systems optimized for tomato to investigate pathogen-associated responses.
The application of genome editing, CRISPR/Cas9 has revolutionized plant breeding by enabling precise, efficient, and targeted modification of native genes, significantly accelerating the development of improved agronomic traits of crops. Therefore, CRISPR/Cas9 technology currently the most extensively used genome editi...
Natural variation in PAM recognition among SaCas9 orthologs is analyzed and StaCas9 is identified as a compact and efficient nuclease recognizing an NNG PAM, establishing StaCas9 as a high-performance genome-editing tool for therapeutic applications.
The CRISPR/Cas9 system is utilized to generate a homozygous Mt4 knockout (Mt4-/-) mouse embryonic stem cell (mESC) line that maintains normal morphology, pluripotency, and the ability to differentiate into all three germ layers.
Traditional methods of diagnosis of TB and mechanisms and challenges related to CRISPR-Cas are included, including the novel technology CRISPR-Cas system for diagnosis, treatment, and the development of new TB vaccines.
This review summarizes CRISPR principles, highlights technical breakthroughs and crop applications that mitigate biotic and abiotic stresses, and outlines practical challenges and future directions necessary for responsible deployment in agriculture.
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