| Literature DB >> 30274789 |
Li Lin1, Xiubin He1, Tianyuan Zhao1, Lingkai Gu1, Yeqing Liu1, Xiaoyu Liu1, Hongyan Liu1, Fayu Yang1, Mengjun Tu1, Lianchao Tang1, Xianglian Ge1, Changbao Liu2, Junzhao Zhao2, Zongming Song1, Jia Qu1, Feng Gu3.
Abstract
FnCpf1-mediated genome-editing technologies have enabled a broad range of research and medical applications. Recently, we reported that FnCpf1 possesses activity in human cells and recognizes a more compatible PAM (protospacer adjacent motif, 5'-KYTV-3'), compared with the other two commonly used Cpf1 enzymes (AsCpf1 and LbCpf1), which requires a 5'-TTTN-3' PAM. However, due to the efficiency and fidelity, FnCpf1-based clinical and basic applications remain a challenge. The direct repeat (DR) sequence is one of the key elements for FnCpf1-mediated genome editing. In principle, its engineering should influence the corresponding genome-editing activity and fidelity. Here we showed that the DR mutants [G(-9)A and U(-7)A] could modulate FnCpf1 performance in human cells, enabling enhancement of both genome-editing efficiency and fidelity. These newly identified features will facilitate the design and optimization of CRISPR-Cpf1-based genome-editing strategies.Entities:
Keywords: FnCpf1; direct repeat; genome editing
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Year: 2018 PMID： 30274789 PMCID： PMC6224799 DOI： 10.1016/j.ymthe.2018.08.021
Source DB: PubMed Journal: Mol Ther ISSN： 1525-0016 Impact factor: 11.454