PNA-Assisted DNAzymes to Cleave Double-Stranded DNA for Genetic Engineering with High Sequence Fidelity.
PNA-Assisted DNAzymes to Cleave Double-Stranded DNA for Genetic Engineering with High Sequence Fidelity.
复制标题
pna辅助DNAzymes在高序列保真度的基因工程中切割双链DNA。
DOI：
10.1021/jacs.1c03129
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发表时间：
2021-07-07
影响因子：
15
通讯作者：
Lu, Yi
中科院分区：
文献类型：
--
作者：
Lyu, Mingkuan;Kong, Linggen;Yang, Zhenglin;Wu, Yuting;McGhee, Claire E.;Lu, Yi
DNAzymes自1994年被发现以来，已广泛应用于传感和成像领域，但由于其底物范围主要局限于单链DNA或RNA，而遗传信息主要存储在双链DNA （dsDNA）中，因此很少用于基因工程。为了克服这一主要限制，我们在此报道了DNAzymes （PANDA）的肽核酸（PNA）辅助双链DNA切割（PANDA）作为将DNAzyme活性扩展到dsDNA的第一个例子。我们发现PANDA是可编程的，可以有效地在目标dsDNA上切割或引起双链断裂，它模仿蛋白质核酸酶，可以在分子克隆中充当限制性内切酶。除了比蛋白酶小得多之外，在我们测试的条件下，PANDA与CRISPR/Cas相比具有更高的序列保真度，这表明它具有作为基因工程和其他生化应用的新型替代工具的潜力。
DNAzymes have been widely used in many sensing and imaging applications but have rarely been used for genetic engineering since their discovery in 1994, because their substrate scope is mostly limited to single-stranded DNA or RNA, whereas genetic information is stored mostly in double-stranded DNA (dsDNA). To overcome this major limitation, we herein report peptide nucleic acid (PNA)-assisted double-stranded DNA nicking by DNAzymes (PANDA) as the first example to expand DNAzyme activity toward dsDNA. We show that PANDA is programmable in efficiently nicking or causing double strand breaks on target dsDNA, which mimics protein nucleases and can act as restriction enzymes in molecular cloning. In addition to being much smaller than protein enzymes, PANDA has a higher sequence fidelity compared with CRISPR/Cas under the condition we tested, demonstrating its potential as a novel alternative tool for genetic engineering and other biochemical applications.
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DOI：
10.1038/mtna.2016.48
发表时间：
2016-07-12
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
Glaser A;McColl B;Vadolas J
通讯作者：
Vadolas J
影响因子：
3.2
作者：
Bahal, Raman;Sahu, Bichismita;Rapireddy, Srinivas;Lee, Chong-Min;Ly, Danith H.
通讯作者：
Ly, Danith H.
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
15
作者：
He, Gaofei;Rapireddy, Sirinivas;Ly, Danith H.
通讯作者：
Ly, Danith H.
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle