Cas9/gRNA-mediated genome editing of yeast mitochondria and Chlamydomonas chloroplasts.

ABSTRACT: We present a new approach to edit both mitochondrial and chloroplast genomes. Organelles have been considered off-limits to CRISPR due to their impermeability to most RNA and DNA. This has prevented applications of Cas9/gRNA-mediated genome editing in organelles while the tool has been widely used for engineering of nuclear DNA in a number of organisms in the last several years. To overcome the hurdle, we designed a new approach to enable organelle genome editing. The plasmids, designated "Edit Plasmids," were constructed with two expression cassettes, one for the expression of Cas9, codon-optimized for each organelle, under promoters specific to each organelle, and the other cassette for the expression of guide RNAs under another set of promoters specific to each organelle. In addition, E

SUBMITTER: Yoo BC 

PROVIDER: S-EPMC6951285 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature