Prokaryote-derived Mbp_Argonaute protein and application thereof

A prokaryotic and protein technology, applied in the field of molecular biology, can solve the problems of time-consuming, no targeted cleavage, and high price of chemically synthesized dsRNA
CN113373129AActive Publication Date: 2021-09-10HUBEI UNIV
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
HUBEI UNIV
Publication Date
2021-09-10

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Abstract

The invention discloses a prokaryote-derived Mbp_Argonaute protein and application thereof. The Mbp_Argonaute protein has an amino acid sequence shown in SEQ ID NO: 1 or as shown in a sequence which has at least 50% or at least 80% of homology with SEQ ID NO: 1. According to the invention, an Argonaute protein gene derived from a cold-resistant prokaryotic organism Mucilaginibacter paaluis is synthesized, the protein is named as MbpAgo, and researches find that the MbpAgo has binding activity to single-stranded guide DNA and has nuclease activity to target RNA and / or target DNA complementarily paired with the single-stranded guide DNA, therefore, the MbpAgo can be used for targeted RNA editing in vivo and in vitro, and then specific site modification of genetic material. The MbpAgo not only can modify RNA with a high-grade structure, but also does not influence an endogenous RNAi pathway of animal and plant cells, provides a brand new powerful tool for RNA editing, and is high in cutting activity and good in specificity.
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Description

technical field

[0001] The invention belongs to the technical field of molecular biology, and in particular relates to a prokaryote-derived Mbp_Argonaute protein and an application thereof. Background technique

[0002] At present, Argonaute protein (eAgos for short) derived from eukaryotes can catalyze the RNA cleavage reaction guided by RNA guide (gRNA) at room temperature, and play a vital role in the RNA interference (RNA interference, RNAi) pathway in vivo . Prokaryotic-derived Argonaute proteins (referred to as pAgos) are more diverse in function and structure than eAgos, but their physiological functions have long been elusive. Early studies mainly focused on pAgos from high-temperature biological sources, except that MpAgo from Marinitoga piezophila prefers to use 5' terminal hydroxylated (5'OH) guide RNA (gRNA) to cleave target single-stranded DNA (single-stranded DNA, ssDNA) and target Except for RNA, pAgos from other high-temperature sources prefer to use gDNA p...

Claims

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