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

PeerJ. 2020; 
Yoo BC, Yadav NS, Orozco EM Jr, Sakai H.
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Gene Synthesis … psbA 3′ UTR. The expression cassette was synthesized in three parts by GenScript. Synthesized DNA … rbcL 3′ UTR. The components were synthesized at GenScript, assembled using BsaI and ligated into YP4. The resulting … Get A Quote

摘要

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 guid... More

关键词

CRISPR; Cas9/gRNA; Chlamydomonas; Chloroplast; DNA replacement; Edit Plasmid; Genome editing; Mitochondria; Organelle; Yeast
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