Genotyping genome-edited mutations in plants using CRISPR ribonucleoprotein complexes.

ABSTRACT: Despite the great achievements in genome editing, accurately detecting mutations induced by sequence-specific nucleases is still a challenge in plants, especially in polyploidy plants. An efficient detection method is particularly vital when the mutation frequency is low or when a large population needs to be screened. Here, we applied purified CRISPR ribonucleoprotein complexes to cleave PCR products for genome-edited mutation detection in hexaploid wheat and diploid rice. We show that this mutation detection method is more sensitive than Sanger sequencing and more applicable than PCR/RE method without the requirement for restriction enzyme site. We also demonstrate that this detection method is especially useful for genome editing in wheat, because target sites are often surrounded by si

SUBMITTER: Liang Z 

PROVIDER: S-EPMC6230946 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature