Tran, Binh Thanh Go, Seulgi Kim, Kyoung-Ran Chung, Hak Suk Ahn, Dae-Ro 2024-07-11T06:30:31Z 2024-07-11T06:30:31Z 2024-07-11 2024-06 2046-2069 https://pubs.kist.re.kr/handle/201004/150220 Post-transcriptional modifications on the guide RNAs utilized in the Cas9 system may have the potential to impact the activity of Cas9. In this study, we synthesized a series of tracrRNAs containing N-6-methyadenosine (m6A), a prevalent post-transcriptional modification, at various positions. We evaluated the effect of these modifications on the DNA cleavage activity of Cas9. Our results show that multiple m6As in the anti-repeat region of tracrRNA reduce the DNA cleavage activity of Cas9. This suggests that the m6A-modified tracrRNA can be used for Cas9 only when the number and the position of the modified residue are properly chosen in tracrRNA. English Royal Society of Chemistry The positional and numerical effect of N6-methyladenosine in tracrRNA on the DNA cleavage activity of Cas9 Article 10.1039/d4ra03957b 1 RSC Advances, v.14, no.29, pp.20529 - 20535 RSC Advances 14 29 20529 20535 Y scie scopus 001256390900001 Chemistry, Multidisciplinary Chemistry Article