Chromosomal Engineering through CRISPR– Cas Technology: A Way Forward
Rao Saad Rehman *
College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
Asad Nadeem Pasha
Department of Plant Pathology, Bahauddin Zakariya University, Multan, Pakistan.
Syed Ali Zafar
Oilseeds Research Institute, Ayub Agricultural Research Institute, Faisalabad, Pakistan.
Mujahid Ali
Department of Plant Breeding and Genetics, Nanjing Agricultural University, Nanjing, China.
Muhammad Waseem
Department of Plant Breeding and Genetics, Bahauddin Zakariya University, Multan, Pakistan.
Muhammad Ahmad
Department of Plant Breeding and Genetics, Nanjing Agricultural University, Nanjing, China.
Nabi Ahmad
Department of Seed Science, University of Agriculture Faisalabad, Pakistan.
Ameer Hamza Hafeez
Department of Horticulture, Bahauddin Zakariya University, Multan, Pakistan.
*Author to whom correspondence should be addressed.
Abstract
The breeding of crops is dependent on the potential to interrupt or maintain genetic links between characteristics, and the availability of genetic variability. CRISPR-Cas is a new genome-editing technique that has made it possible for breeders to introduce regulated and site-specific genetic diversity while simultaneously improving qualities with high efficacy. The existence of genomic linkage is a barrier in transferring desirable features among domesticated species from their wild counterparts. One way to address this issue is to create mutants with deficiencies in the meiotic recombination machinery, thereby enhancing global crossover frequencies between homologous parental chromosomes. Although this seemed to be a promising approach at first, thus far, no crossover frequencies could be enhanced in recombination-cold regions of the genome. Consequently, attempts have been made to induce site-specific DSBs in both somatic and meiotic plant cells by utilizing CRISPR–Cas techniques to achieve preset crossovers among homologs. Nonetheless, this method has not yielded significant heritable homologous crossings which were recombination-based. Lately, CRISPR–Cas has been used to achieve hereditary chromosomal rearrangements (CRs), including translocations and inversions, in plants. This method allows for the development of megabase CRs by DSB repair through non-homologous end-joining after insertion of DSBs in somatic plant cells. This technique may potentially make it possible to restructure genomes on a more global scale, culminating in the creation not just of synthetic plant chromosomes, but also that of new plant species.
Keywords: CRISPR-Cas, genetic linkage, crossover induction, chromosome engineering, reciprocal translocations, synthetic chromosomes, CRISPR-associated protein