Open AccessUnexpected binding behaviors of bacterial Argonautes in human cells cast doubts on their use as targetable gene regulators
Author(s) -
Henriette O’Geen,
Chonghua Ren,
Nicole B. Coggins,
Sofie Bates,
David J. Segal
Publication year - 2018
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0193818
Subject(s) - argonaute , biology , crispr , genome editing , dna , epigenetics , gene , computational biology , epigenome , cas9 , genetics , dna methylation , microbiology and biotechnology , gene expression , rna interference , rna
Prokaryotic Argonaute proteins (pAgos) have been proposed as an alternative to the CRISPR/Cas9 platform for gene editing. Although Argonaute from Natronobacterium gregoryi ( Ng Ago) was recently shown unable to cleave genomic DNA in mammalian cells, the utility of Ng Ago or other pAgos as a targetable DNA-binding platform for epigenetic editing has not been explored. In this report, we evaluated the utility of two prokaryotic Argonautes ( Ng Ago and Tt Ago) as DNA-guided DNA-binding proteins. Ng Ago showed no meaningful binding to chromosomal targets, while Tt Ago displayed seemingly non-specific binding to chromosomal DNA even in the absence of guide DNA. The observed lack of DNA-guided targeting and unexpected guide-independent genome sampling under the conditions in this study provide evidence that these pAgos might be suitable for neither gene nor epigenome editing in mammalian cells.