Natronobacterium gregoryi sp2 cells">Characterization of endogenous nucleic acids that bind to NgAgo in Natronobacterium gregoryi sp2 cells

[摘要] As nucleic acid-guided endonucleases, some prokaryotic Argonautes have been used as programmablenucleases. Natronobacterium gregoryi Argonaute (NgAgo) has also been proposed for gene editing, but this remainsvery controversial. Until now, the endogenous nucleic acids that bind to NgAgo in Natronobacterium gregoryi sp2(N. gregoryi sp2) have not been characterized. We expressed the conserved PIWI domain of NgAgo and used it toinduce anti-PIWI antibody. We also cultured the N. gregoryi sp2 strain and performed immunoprecipitation,chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RIP) assays. The nucleic acids thatendogenously bound NgAgo in N. gregoryi sp2 cells were sequenced and analyzed. The results showed that NgAgoendogenously bound RNA rather than DNA. NgAgo-associated RNAs were mainly transcripts of genes that encodedtRNA, transcriptional regulators, RNA polymerases, and RNA-binding proteins. NgAgo mainly binds to thetranscripts inside genes or in their upstream sequences. Interestingly, the top enriched motif of peaks was the same asthat of miR-1289, suggesting that NgAgo may regulate gene expression post-transcriptionally. GO enrichment analysisshowed that the peak-associated genes were enriched in transmembrane transport processes. These results revealedthat NgAgo binds RNA and may function in post-transcriptional regulation in vivo.

[学科分类] 仪器

[关键词] NgAgo;PIWI;Immunoprecipitation;ChIP-seq;RIP-seq;Post-transcriptional regulation