biostudies-literatureRamadan MNational Key Research and Development Program of China16https://www.ebi.ac.uk/biostudies/studies/S-EPMC7869495biostudies-literatureUnknown17(1)<h4>Background</h4>Upland cotton (Gossypium hirsutum), harboring a complex allotetraploid genome, consists of A and D sub-genomes. Every gene has multiple copies with high sequence similarity that makes genetic, genomic and functional analyses extremely challenging. The recent accessibility of CRISPR/Cas9 tool provides the ability to modify targeted locus efficiently in various complicated plant genomes. However, current cotton transformation method targeting one gene requires a complicated, long and laborious regeneration process. Hence, optimizing strategy that targeting multiple genes is of great value in cotton functional genomics and genetic engineering.<h4>Results</h4>To target multiple genes in a single experiment, 112 plant development-related genes were knocked out via optimized CPlant methodsEfficient CRISPR/Cas9 mediated Pooled-sgRNAs assembly accelerates targeting multiple genes related to male sterility in cotton.PMC78694952016YFD0101402Min LRamadan MAlariqi MLi YLiu ZJin SZhang XMa YZhang RfalseEfficient CRISPR/Cas9 mediated Pooled-sgRNAs assembly accelerates targeting multiple genes related to male sterility in cotton.<h4>Background</h4>Upland cotton (Gossypium hirsutum), harboring a complex allotetraploid genome, consists of A and D sub-genomes. Every gene has multiple copies with high sequence similarity that makes genetic, genomic and functional analyses extremely challenging. The recent accessibility of CRISPR/Cas9 tool provides the ability to modify targeted locus efficiently in various complicated plant genomes. However, current cotton transformation method targeting one gene requires a complicated, long and laborious regeneration process. Hence, optimizing strategy that targeting multiple genes is of great value in cotton functional genomics and genetic engineering.<h4>Results</h4>To target multiple genes in a single experiment, 112 plant development-related genes were knocked out via optimized C2021-01-01T00:00:00Z2021 Feb2025-04-18T15:58:12.194Z2025-04-07T02:53:37.688ZS-EPMC78694953355788910.1186/s13007-021-00712-x