Breakthrough in CRISPR/Cas system: Current and future directions and challenges
Ahmad Ali, Muhammad Mubashar Zafar, Zunaira Farooq, Syed Riaz Ahmed, Aqsa Ijaz, Zunaira Anwar, Huma Abbas, Muhammad Sayyam Tariq, Hala Tariq, Mahwish Mustafa, M Bajwa, Fiza Shaukat, Abdul Razzaq, Ren Maozhi
Abstract
Targeted genome editing (GE) technology has brought a significant revolution in fictional genomic research and given hope to plant scientists to develop desirable varieties. This technology involves inducing site-specific DNA perturbations that can be repaired through DNA repair pathways. GE products currently include CRISPR-associated nuclease DNA breaks, prime editors generated DNA flaps, single nucleotide-modifications, transposases, and recombinases. The discovery of double-strand breaks, site-specific nucleases (SSNs), and repair mechanisms paved the way for targeted GE, and the first-generation GE tools, ZFNs and TALENs, were successfully utilized in plant GE. However, CRISPR-Cas has now become the preferred tool for GE due to its speed, reliability, and cost-effectiveness. Plant functional genomics has benefited significantly from the widespread use of CRISPR technology for advancements and developments. This review highlights the progress made in CRISPR technology, including multiplex editing, base editing (BE), and prime editing (PE), as well as the challenges and potential delivery mechanisms.