Evaluation of CRISPR/Cas9 Constructs in Wheat Cell Suspension Cultures.

ABSTRACT: Despite intensive optimization efforts, developing an efficient sequence-specific CRISPR/Cas-mediated genome editing method remains a challenge, especially in polyploid cereal species such as wheat. Validating the efficacy of nuclease constructs prior to using them in planta is, thus, a major step of every editing experiment. Several construct evaluation strategies were proposed, with PEG-mediated plasmid transfection of seedling-derived protoplasts becoming the most popular. However, the usefulness of this approach is affected by associated construct copy number bias and chromatin relaxation, both influencing the outcome. Therefore, to achieve a reliable evaluation of CRISPR/Cas9 constructs, we proposed a system based on an Agrobacterium-mediated transformation of established wheat

SUBMITTER: Michalski K 

PROVIDER: S-EPMC9916915 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature