The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in le...The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in lepidopterans has not yet been reported.In the present study,we evaluated the RNA cleavage activity of the CRISPR-Cas13 system in the silkworm(Bombyx mori),a model lepidopteran insect,both ex vivo and in vivo.We established two stable silkworm BmE cell lines expressing PspCas13b and CasRx,respectively.Further analysis demonstrated that both PspCas13b and CasRx effectively down-regulated the transcription of exogenouslyintroduced target and endogenous genes in these cell lines.In addition,we generated two transgenic silkworm strains,one expressing CasRx and the other expressing RNA-guided CRISPR RNA targeting Sex combs reduced(Scr).Further crossing experiments showed that CasRx induced a down-regulation of Scr transcription in silkworms,which impaired systemic growth of larvae.Overall,this study demonstrated that the CRISPR-Cas13RNA editing system works efficiently in the silkworm,providing a potential alternative approach for RNA manipulation in lepidopteran insects.展开更多
A paper titled"The CRISPR-Cas13 a gene-editing system induces collateral cleavage of RNA in glioma cells",recently published in Advanced Science by the Kang group,reports the promising application of the CRI...A paper titled"The CRISPR-Cas13 a gene-editing system induces collateral cleavage of RNA in glioma cells",recently published in Advanced Science by the Kang group,reports the promising application of the CRISPR-Cas13 a system in cancer cells1.展开更多
CRISPR-Cas13系统(clustered regularly interspaced short palindromic repeats associated Cas system,CRISPR-Cas)是一种快速、高效、精准的新型RNA编辑工具,具有易于设计、结构简单、操作方便、特异性强的特点。综述了CRISPR-Cas13...CRISPR-Cas13系统(clustered regularly interspaced short palindromic repeats associated Cas system,CRISPR-Cas)是一种快速、高效、精准的新型RNA编辑工具,具有易于设计、结构简单、操作方便、特异性强的特点。综述了CRISPR-Cas13在CRISPR分类系统中的地位、CRISPR-Cas13的结构基础以及作用机制、与其他RNA水平调节方法的比较以及目前的应用前景,以期为相关研究提供参考。展开更多
Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose postt...Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose posttranscriptional regulations such as alternative splicing,alternative polyadenylation and modifications impact its coded protein expression and functions.However,the functional significance and therapeutic potential of RNA posttranscriptional regulations are not well studied due to the lack of suitable RNA engineering platforms.The discovery of a novel CRISPR-Cas system termed CRISPR-Cas13 in 2015 that specifically targets RNA templates brought a new role to CRISPR to target and edit RNA with high specificity,which opened a new era of RNA manipulations to some degree.This review will summarize the emerging applications of the catalytically inactive CRISPR-Cas13 system(CRISPR-dCas13)in mRNA engineering and highlight the prospection of the CRISPR-dCas13 system for other RNA modification regulations and its therapeutic potential.展开更多
基金supported by the National Natural Science Foundation of China(32070496,32370555)Fundamental Research Funds for the Central Universities(SWU120033)Technology Innovation and Application Development Program of Chongqing(CSTB2024TIADKPX0023)。
文摘The CRISPR-Cas13 system,an RNA-guided editing tool,has emerged as a highly efficient and stable RNA editing technique.Although the CRISPR-Cas13 system has been developed in several insect species,its application in lepidopterans has not yet been reported.In the present study,we evaluated the RNA cleavage activity of the CRISPR-Cas13 system in the silkworm(Bombyx mori),a model lepidopteran insect,both ex vivo and in vivo.We established two stable silkworm BmE cell lines expressing PspCas13b and CasRx,respectively.Further analysis demonstrated that both PspCas13b and CasRx effectively down-regulated the transcription of exogenouslyintroduced target and endogenous genes in these cell lines.In addition,we generated two transgenic silkworm strains,one expressing CasRx and the other expressing RNA-guided CRISPR RNA targeting Sex combs reduced(Scr).Further crossing experiments showed that CasRx induced a down-regulation of Scr transcription in silkworms,which impaired systemic growth of larvae.Overall,this study demonstrated that the CRISPR-Cas13RNA editing system works efficiently in the silkworm,providing a potential alternative approach for RNA manipulation in lepidopteran insects.
基金supported by grants from the National Natural Science Foundation of China(Grant No.81772682,81974389)Jiangsu Province’s Key Discipline of Medicine(Grant No.ZDXKA2016001)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘A paper titled"The CRISPR-Cas13 a gene-editing system induces collateral cleavage of RNA in glioma cells",recently published in Advanced Science by the Kang group,reports the promising application of the CRISPR-Cas13 a system in cancer cells1.
文摘CRISPR-Cas13系统(clustered regularly interspaced short palindromic repeats associated Cas system,CRISPR-Cas)是一种快速、高效、精准的新型RNA编辑工具,具有易于设计、结构简单、操作方便、特异性强的特点。综述了CRISPR-Cas13在CRISPR分类系统中的地位、CRISPR-Cas13的结构基础以及作用机制、与其他RNA水平调节方法的比较以及目前的应用前景,以期为相关研究提供参考。
文摘Posttranscriptional regulations of different types of RNA,including rRNA,tRNA,mRNA and ncRNA are widely involved in normal physiology and diseases.m RNA,as the intermediary product between gene and protein,whose posttranscriptional regulations such as alternative splicing,alternative polyadenylation and modifications impact its coded protein expression and functions.However,the functional significance and therapeutic potential of RNA posttranscriptional regulations are not well studied due to the lack of suitable RNA engineering platforms.The discovery of a novel CRISPR-Cas system termed CRISPR-Cas13 in 2015 that specifically targets RNA templates brought a new role to CRISPR to target and edit RNA with high specificity,which opened a new era of RNA manipulations to some degree.This review will summarize the emerging applications of the catalytically inactive CRISPR-Cas13 system(CRISPR-dCas13)in mRNA engineering and highlight the prospection of the CRISPR-dCas13 system for other RNA modification regulations and its therapeutic potential.
