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A programmable pAgo nuclease with universal guide and target specificity from the mesophilic bacterium Kurthia massiliensisстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 17 июня 2021 г.
- Авторы: Kropocheva Ekaterina, Kuzmenko Anton, Aravin Alexei A., Esyunina Daria, Kulbachinskiy Andrey
- Журнал: Nucleic Acids Research
- Том: 49
- Номер: 7
- Год издания: 2021
- Издательство: Oxford University Press
- Местоположение издательства: United Kingdom
- Первая страница: 4054
- Последняя страница: 4065
- DOI: 10.1093/nar/gkab182
- Аннотация: Argonaute proteins are programmable nucleases that are found in both eukaryotes and prokaryotes and provide defense against invading genetic elements. Although some prokaryotic argonautes (pAgos) were shown to recognize RNA targets in vitro, the majority of studied pAgos have strict specificity toward DNA, which limits their practical use in RNA-centric applications. Here, we describe a unique pAgo nuclease, KmAgo, from the mesophilic bacterium Kurthia massiliensis that can be programmed with either DNA or RNA guides and can precisely cleave both DNA and RNA targets. KmAgo binds 16-20 nt long 5′-phosphorylated guide molecules with no strict specificity for their sequence and is active in a wide range of temperatures. In bacterial cells, KmAgo is loaded with small DNAs with no obvious sequence preferences suggesting that it can uniformly target genomic sequences. Mismatches between the guide and target sequences greatly affect the efficiency and precision of target cleavage, depending on the mismatch position and the nature of the reacting nucleic acids. Target RNA cleavage by KmAgo depends on the formation of secondary structure indicating that KmAgo can be used for structural probing of RNA. These properties of KmAgo open the way for its use for highly specific nucleic acid detection and cleavage.
- Добавил в систему: Каменский Петр Андреевич