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Ago nucleases from Clostridium butyricum and Limnothrix rosea can process DNA substrates at moderate temperatures

Kuzmenko, A. and Yudin, D. and Aravin, A. and Kulbachinskiy, A. (2019) Ago nucleases from Clostridium butyricum and Limnothrix rosea can process DNA substrates at moderate temperatures. FEBS Open Bio, 9 (S1). p. 400. ISSN 2211-5463. https://resolver.caltech.edu/CaltechAUTHORS:20191017-154306053

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Abstract

Prokaryotic Argonaute proteins (pAgos) are diverse homologs of eukaryotic Argonautes (eAgos) involved in RNA interference. In contrast to eAgos, which are RNA-guided RNA nucleases, several pAgos were reported to act as DNA nucleases suggesting that they may be used as an alternative to CRISPRC as nucleases for genome editing. However, all previously studied pAgos were isolated from thermophilic bacteria or archaea thus limiting their potential use in genomic applications. We describe two pAgo nucleases from mesophilic bacteria, Clostridium butyricum (CbAgo) and Limnothrix rosea (LrAgo). Both CbAgo and LrAgo use small DNA guides to cleave complementary DNA targets and are active at physiological temperatures. At the same time, the two proteins reveal significant variations in DNA processing depending on the reaction conditions and guide structure, including changes in the guide 5’-end and the presence of mismatches. We show that CbAgo is highly active under a wide range of conditions and can precisely cleave single-stranded and double-stranded DNA at moderate temperatures suggesting that it may be used for DNA manipulations both in vitro and in vivo.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/2211-5463.12675DOIAbstracts
ORCID:
AuthorORCID
Kulbachinskiy, A.0000-0002-2292-6424
Additional Information:© 2019 The Authors. FEBS Open Bio © 2019 FEBS. Issue Online: 04 July 2019. This work was supported by the grant 14.W03.31.0007 of the Ministry of Science and Higher Education of the Russian Federation.
Funders:
Funding AgencyGrant Number
Ministry of Education and Science of the Russian Federation14.W03.31.0007
Issue or Number:S1
Record Number:CaltechAUTHORS:20191017-154306053
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20191017-154306053
Official Citation:(2019), Posters. FEBS Open Bio, 9: 65-431. doi:10.1002/2211-5463.12675
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:99342
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:19 Oct 2019 04:29
Last Modified:09 Mar 2020 13:18

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