核糖核酸
生物发生
生物
核酸酶
转座因子
遗传学
DNA
核糖核蛋白
计算生物学
非编码RNA
细胞生物学
基因组
基因
作者
Suchita P. Nety,Han Altae-Tran,Soumya Kannan,F. Esra Demircioglu,Guilhem Faure,Seiichi Hirano,Kepler Mears,Yugang Zhang,Rhiannon K. Macrae,Feng Zhang
出处
期刊:The CRISPR journal
[Mary Ann Liebert]
日期:2023-06-01
卷期号:6 (3): 232-242
被引量:9
标识
DOI:10.1089/crispr.2023.0015
摘要
TnpB is a member of the Obligate Mobile Element Guided Activity (OMEGA) RNA-guided nuclease family, is harbored in transposons, and likely functions to maintain the transposon in genomes. Previously, it was shown that TnpB cleaves double- and single-stranded DNA substrates in an RNA-guided manner, but the biogenesis of the TnpB ribonucleoprotein (RNP) complex is unknown. Using in vitro purified apo TnpB, we demonstrate the ability of TnpB to generate guide omegaRNA (ωRNA) from its own mRNA through 5' processing. We also uncover a potential cis-regulatory mechanism whereby a region of the TnpB mRNA inhibits DNA cleavage by the RNP complex. We further expand the characterization of TnpB by examining ωRNA processing and RNA-guided nuclease activity in 59 orthologs spanning the natural diversity of the TnpB family. This work reveals a new functionality, ωRNA biogenesis, of TnpB, and characterizes additional members of this biotechnologically useful family of programmable enzymes.
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