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- Title
Chimeric crRNAs Retaining Activity of Cas12a with Potential to Improve Specificity.
- Authors
Park, Jihyun; Choi, Jaewoo; Duong, Men Thi Hoai; Ahn, Hee‐Chul; Hong, Seung Woo; Hwang, Gil Tae; An, Jinsu; Chung, Hak Suk; Ahn, Dae‐Ro
- Abstract
Keywords: CRISPR-Cas; Cas12a; Chimeric crRNA EN CRISPR-Cas Cas12a Chimeric crRNA 87 90 4 01/29/21 20210101 NES 210101 RNA residues in the guide region of crRNA for Cas12a can be partially replaced by DNA. (d) DNA cleavage reaction rate (kobs) (right) of Cas12a guided by chimeric crRNAs containing various numbers of DNA residues (left). gl Recently, the effect of DNA sugar backbone in guide RNAs of Cas9 has been investigated. In contrast, the chimeric crRNAs containing nine or more DNA residues (9 DNA, 10 DNA, 11 DNA, and 14 DNA) showed substantially lowered rate constants (0.36-0.09 min SP -1 sp ). (c) Melting temperatures (Tm) of 21-mer RNA/DNA duplexes with various numbers of DNA replacement in the RNA strand. gl We next examined whether the cleavage activity could be dependent on the thermal stability of the crRNA/target-DNA duplex.
- Subjects
NUCLEIC acids; HELICAL structure; MICROBIAL genes; NUCLEOTIDE sequence
- Publication
Bulletin of the Korean Chemical Society, 2021, Vol 42, Issue 1, p87
- ISSN
0253-2964
- Publication type
Article
- DOI
10.1002/bkcs.12109