在Gluconobacter oxydans中的抗CRISPR蛋白通过乙化使FnCas12a失活
Xuyang Wang1, Dong Li2, Zhijie Qin2
1Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
International journal of biological macromolecules
|January 10, 2025
概括
研究人员在Gluconobacter oxidans中发现了抗CRISPR蛋白AcrVA6,AcrVA7和AcrVA8,这些蛋白通过乙化使FnCas12a失活. 基因组脱乙酶HDAC11可以逆转这种情况,从而实现可逆基因编辑控制.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 葡萄糖菌氧化剂是维生素C生产的关键宿主.
- 克里斯普尔/Cas12a系统对于基因编辑至关重要,但在G. oxydans.中自然是不活跃的.
- 在G. oxydans中CRISPR/Cas12a失活的精确机制以前是未知的.
研究的目的:
- 为了阐明G. oxydans.中CRISPR/Cas12a失活的机制.
- 确定特定的抗CRISPR蛋白及其作用模式.
- 通过乙化探索可逆基因编辑调节的潜力.
主要方法:
- 对抗CRISPR蛋白的鉴定和特征 (AcrVA6,AcrVA7,AcrVA8) 的研究.
- 生物化学分析以确定FnCas12a无活化的乙化位点和机制.
- 量子力学计算来分析乙化途径.
- 调查基因素脱乙酶 (HDAC11) 的参与.
主要成果:
- 通过向特定的氨酸残留物 (Lys671,Lys589,Lys823) 鉴定出AcrVA6,AcrVA7和AcrVA8是通过向特定的氨酸残留物 (Lys671,Lys589,Lys823) 来非激活FnCas12a的乙转移酶.
- AcrVA6和AcrVA8破坏了DNA结合,而AcrVA7则抑制了复合体的形成.
- 发现HDAC11可以逆转AcrVA6介导的乙化.
- AcrVA6和HDAC11作为可逆FnCas12a控制的可光开关调节器.
结论:
- 这项研究揭示了由特定的抗CRISPR蛋白质对Cas12a无活化的基于乙化的多种机制.
- 由AcrVA6和HDAC11调节的可逆乙化提供了一种用于控制基因编辑工具的新方法.
- 这些发现为乙化在基因编辑调节中的作用及其潜在应用提供了新的见解.
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