生物技术通报2025，Vol.41Issue(3)：71-82,12.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0750
非编码RNA的引导RNA的设计及其应用
Design and Application of Guide RNAs for Non-coding RNAs
摘要
Abstract
Non-coding RNA(ncRNA)refers to functional RNA molecules that typically do not encode proteins but play important physiological roles by regulating the expression and function of coding genes.microRNA(miRNA),circular RNA(circRNA),and long non-coding RNA(lncRNA)are currently known as the three major types of regulatory ncRNAs with important physiological and pathological significance.The loss,rearrangement,and dysregulation of their genes are closely related to the occurrence and development of various diseases.Clustered regularly interspaced short palindromic repeats(CRISPR)and CRISPR-associated proteins(CRISPR/Cas)are types of immune-defense systems widely existing in bacteria and archaea,consisting of CRISPR-associated proteins(Cas)and guide RNAs(gRNAs).Cas nucleases shear and remove exogenous genes,while gRNAs determine their precise recognition and effective cleavage of target DNA/RNAs.CRISPR/Cas technology has excellent capabilities of site-specific genome editing and cleavage of RNA transcripts and can precisely and efficiently edit,intervene,and detect abnormalities in these ncRNAs and thus contribute to successful diagnosis and treatment of diseases.This paper summarizes the composition and design principles of Cas9 and Cas13 gRNA and the unique molecular characteristics of miRNA,circRNA,and lncRNA genes and their respective molecular features produced by the RNA itself or during RNA formation,emphasizes the design strategies and target selection of their gRNAs,as well as how Cas9 and Cas13 nucleases can accurately detect and intervene in abnormal miRNA,circRNA,and lncRNA under the guidance of gRNA with high specificity and efficiency.The article further discusses the similarities and differences in gRNA design among three types of ncRNAs,as well as the challenges faced by CRISPR/Cas applications in ncRNAs.It also gives prospects for future gRNA designs for more accurate targeting at ncRNAs,aiming to provide some reference for the employment of CRISPR/Cas technology in managing ncRNAs-associated diseases.关键词
CRISPR/Cas/guide RNA/非编码RNA/miRNAs/circRNAs/lncRNAs/敲除靶点Key words
CRISPR/Cas/guide RNA/non-coding RNA/miRNAs/circRNAs/lncRNAs/knockdown targets引用本文复制引用
秦玉婷,潘森涛,陈渝萍..非编码RNA的引导RNA的设计及其应用[J].生物技术通报,2025,41(3):71-82,12.基金项目
湖南省自然科学基金项目(2024JJ5342) （2024JJ5342）
