Characterization of endogenous nucleic acids that bind to NgAgo in Natronobacterium gregoryi sp2 cells
Characterization of endogenous nucleic acids that bind to NgAgo in Natronobacterium gregoryi sp2 cells
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格雷戈里嗜盐杆菌 sp2 细胞中与 NgAgo 结合的内源核酸的表征
DOI：
10.32604/biocell.2022.016500
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发表时间：
2022
期刊：
影响因子：
1.2
通讯作者：
Jian Huang
中科院分区：
文献类型：
--
作者：
Lixu Jiang;Lin Ning;Chunchao Pu;Zixin Wang;Bifang He;Jian Huang
As nucleic acid-guided endonucleases, some prokaryotic Argonautes have been used as programmable nucleases. Natronobacterium gregoryi 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 Natronobacterium gregoryi sp2 (N. gregoryi 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 N. gregoryi sp2 strain and performed immunoprecipitation, chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RIP) assays. The nucleic acids that endogenously bound NgAgo in N. gregoryi 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 in vivo.
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通讯作者：
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