Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish
Chong Zhang, Tong Lü, Yizhuang Zhang, Jiaguang Li, Imran Tarique, Fenfen Wen, Aijun Chen, Jia-Sheng Wang, Zhuoyu Zhang, Yanjun Zhang, De‐Li Shi, Ming Shao
Abstract
offspring. Notably, most of these maternal mutants displayed either sgRNA site-spanning genomic deletions or unintended large deletions extending distantly from the sgRNA targets, suggesting a prominent deletion-prone tendency of genome editing in the oocyte. Thus, our method allows maternal gene knockout in the absence of viable and fertile homozygous mutant adults. This approach is particularly time-saving and can be applied for functional screening of maternal factors and generating genomic deletions in zebrafish.
Topics & Concepts
ZebrafishCRISPRTransgeneCas9MutantCell biologyOocyteBiologyGenome editingComputational biologyGeneticsGeneEmbryoCRISPR and Genetic EngineeringAnimal Genetics and ReproductionRNA Interference and Gene Delivery