Unexpected binding behaviors of bacterial Argonautes in human cells cast doubts
on their use as targetable gene regulators
#MMPMID29584750
O'Geen H
; Ren C
; Coggins NB
; Bates SL
; Segal DJ
PLoS One
2018[]; 13
(3
): e0193818
PMID29584750
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Prokaryotic Argonaute proteins (pAgos) have been proposed as an alternative to
the CRISPR/Cas9 platform for gene editing. Although Argonaute from
Natronobacterium gregoryi (NgAgo) was recently shown unable to cleave genomic DNA
in mammalian cells, the utility of NgAgo 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 (NgAgo and TtAgo)
as DNA-guided DNA-binding proteins. NgAgo showed no meaningful binding to
chromosomal targets, while TtAgo 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.
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