-甲基腺苷在反式激活crRNA中对Cas9的DNA切割活性的位置和数量效应。
The positional and numerical effect of -methyladenosine in tracrRNA on the DNA cleavage activity of Cas9.
作者信息
Tran Binh Thanh, Go Seulgi, Kim Kyoung-Ran, Chung Hak Suk, Ahn Dae-Ro
机构信息
Chemical and Biological Integrative Research Center, Korea Institute of Science and Technology (KIST) Hwarangno 14-gil 5, Seongbuk-gu Seoul 02792 Republic of Korea
Division of Bio-Medical Science and Technology, KIST School, University of Science and Technology (UST) Seoul 02792 Republic of Korea.
出版信息
RSC Adv. 2024 Jun 27;14(29):20529-20535. doi: 10.1039/d4ra03957b.
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 -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.
在Cas9系统中使用的向导RNA上的转录后修饰可能会影响Cas9的活性。在本研究中，我们合成了一系列在不同位置含有N6-甲基腺苷（m6A，一种普遍的转录后修饰）的反式激活CRISPR RNA（tracrRNA）。我们评估了这些修饰对Cas9 DNA切割活性的影响。我们的结果表明，tracrRNA反重复区域中的多个m6A会降低Cas9的DNA切割活性。这表明，只有当修饰残基的数量和位置在tracrRNA中得到适当选择时，m6A修饰的tracrRNA才能用于Cas9。
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