|
|
| Type |
Symposium |
| Area |
[IBS 심포지엄] Genome Editing and the CRISPR/Cas Revolution (※IBS 유전체 교정 연구단 공동개최) |
| Room No. |
302호 |
| Time |
WED 15:20-: |
| Code |
KCS4-2 |
| Subject |
Genome editing with a small Cas9 orthologue derived from Campylobacter jejuni |
| Authors |
김은지, 김진수1,*
기초과학연구원 유전체교정연구단, Korea
1서울대학교 화학부, Korea
|
| Abstract |
Several CRISPR-Cas9 orthologues have been used for genome editing. Here, we present the smallest Cas9 orthologue characterized thus far, derived from Campylobacter jejuni (CjCas9), for efficient genome editing in human cells and in mice. We determined protospacer-adjacent motif (PAM) sequences recognized by CjCas9 in vitro and optimized single-guide RNA (sgRNA) length in human cells. Digenome-seq, a genome-wide method for assessing off-target effects, showed that CjCas9 was highly specific, cleaving merely several sites in the human or mouse genome in vitro. Because CjCas9 is smaller than Streptococcus pyogenes Cas9 and Staphylococcus aureus Cas9, we were able to package the CjCas9 gene, its sgRNA sequence, and a marker gene in an all-in-one adeno-associated virus (AAV) vector, suggesting that CjCas9 is a promising new option for the AAV mediated gene and cell therapy. |
| E-mail |
eunjk39@gmail.com |
|
119th General Meeting of the KCS