@Article{biocell.2021.016500,
AUTHOR = {LIXU JIANG, LIN NING, CHUNCHAO PU, ZIXIN WANG, BIFANG HE, JIAN HUANG},
TITLE = {Characterization of endogenous nucleic acids that bind to NgAgo in <i>Natronobacterium gregoryi</i> sp2 cells},
JOURNAL = {BIOCELL},
VOLUME = {46},
YEAR = {2022},
NUMBER = {2},
PAGES = {547--557},
URL = {http://www.techscience.com/biocell/v46n2/45106},
ISSN = {1667-5746},
ABSTRACT = {As nucleic acid-guided endonucleases, some prokaryotic Argonautes have been used as programmable
nucleases. <i>Natronobacterium gregoryi</i> Argonaute (NgAgo) has also been proposed for gene editing, but this remains
very controversial. Until now, the endogenous nucleic acids that bind to NgAgo in <i>Natronobacterium gregoryi</i> sp2
(<i>N. gregoryi</i> sp2) have not been characterized. We expressed the conserved PIWI domain of NgAgo and used it to
induce anti-PIWI antibody. We also cultured the <i>N. gregoryi</i> sp2 strain and performed immunoprecipitation,
chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RIP) assays. The nucleic acids that
endogenously bound NgAgo in <i>N. gregoryi</i> sp2 cells were sequenced and analyzed. The results showed that NgAgo
endogenously bound RNA rather than DNA. NgAgo-associated RNAs were mainly transcripts of genes that encoded
tRNA, transcriptional regulators, RNA polymerases, and RNA-binding proteins. NgAgo mainly binds to the
transcripts inside genes or in their upstream sequences. Interestingly, the top enriched motif of peaks was the same as
that of miR-1289, suggesting that NgAgo may regulate gene expression post-transcriptionally. GO enrichment analysis
showed that the peak-associated genes were enriched in transmembrane transport processes. These results revealed
that NgAgo binds RNA and may function in post-transcriptional regulation <i>in vivo</i>.},
DOI = {10.32604/biocell.2021.016500}
}