脱氧核酶
DNA
化学
劈开
计算生物学
清脆的
肽核酸
核酶
核糖核酸酶H
蛋白质工程
核糖核酸
核酸
遗传学
核糖核酸酶P
生物化学
酶
基因
生物
作者
Mingkuan Lyu,Linggen Kong,Zhenglin Yang,Yuting Wu,Claire E. McGhee,Yi Lu
摘要
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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