| Autor: |
Li, Zhifang, Guo, Ruochen, Sun, Xiaozhi, Li, Guoling, Shao, Zhuang, Huo, Xiaona, Yang, Rongrong, Liu, Xinyu, Cao, Xi, Zhang, Hainan, Zhang, Weihong, Zhang, Xiaoyin, Ma, Shuangyu, Zhang, Meiling, Liu, Yuanhua, Yao, Yinan, Shi, Jinqi, Yang, Hui, Hu, Chunyi, Zhou, Yingsi |
| Předmět: |
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| Zdroj: |
Nature Communications; 1/27/2024, Vol. 15 Issue 1, p1-11, 11p |
| Abstrakt: |
Transposon-associated ribonucleoprotein TnpB is known to be the ancestry endonuclease of diverse Cas12 effector proteins from type-V CRISPR system. Given its small size (408 aa), it is of interest to examine whether engineered TnpB could be used for efficient mammalian genome editing. Here, we showed that the gene editing activity of native TnpB from Deinococcus radiodurans (ISDra2 TnpB) in mouse embryos was already higher than previously identified small-sized Cas12f1. Further stepwise engineering of noncoding RNA (ωRNA or reRNA) component of TnpB significantly elevated the nuclease activity of TnpB. Notably, an optimized TnpB-ωRNA system could be efficiently delivered in vivo with single adeno-associated virus (AAV) and corrected the disease phenotype in a tyrosinaemia mouse model. Thus, the engineered miniature TnpB system represents a new addition to the current genome editing toolbox, with the unique feature of the smallest effector size that facilitate efficient AAV delivery for editing of cells and tissues. Miniature gene editing tools are highly desired for efficient in vivo delivery and disease treatment. Here, the authors reported engineering hypercompact TnpB-ωRNA for robust gene editing with minimal off-target effect in cultured cells and use it to rescue fatal genetic liver disease in a tyrosinaemia mouse model. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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