Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection
In cancer diagnosis, diverse microRNAs (miRNAs) are used as biomarkers for carcinogenesis of distinctive human cancers. Thus, the detection of these miRNAs and their quantification are very important in prevention of cancer diseases in human beings. However, efficient RNA detection often requires RT...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/3/2857 |
| _version_ | 1797624265899507712 |
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| author | Zhenxiao Yu Jianan Xu Qunxin She |
| author_facet | Zhenxiao Yu Jianan Xu Qunxin She |
| author_sort | Zhenxiao Yu |
| collection | DOAJ |
| description | In cancer diagnosis, diverse microRNAs (miRNAs) are used as biomarkers for carcinogenesis of distinctive human cancers. Thus, the detection of these miRNAs and their quantification are very important in prevention of cancer diseases in human beings. However, efficient RNA detection often requires RT-PCR, which is very complex for miRNAs. Recently, the development of CRISPR-based nucleic acid detection tools has brought new promises to efficient miRNA detection. Three CRISPR systems can be explored for miRNA detection, including type III, V, and VI, among which type III (CRISPR-Cas10) systems have a unique property as they recognize RNA directly and cleave DNA collaterally. In particular, a unique type III-A Csm system encoded by <i>Lactobacillus delbrueckii</i> subsp. <i>bulgaricus</i> (LdCsm) exhibits robust target RNA-activated DNase activity, which makes it a promising candidate for developing efficient miRNA diagnostic tools. Herein, LdCsm was tested for RNA detection using fluorescence-quenched DNA reporters. We found that the system is capable of specific detection of miR-155, a microRNA implicated in the carcinogenesis of human breast cancer. The RNA detection system was then improved by various approaches including assay conditions and modification of the 5′-repeat tag of LdCsm crRNAs. Due to its robustness, the resulting LdCsm detection platform has the potential to be further developed as a better point-of-care miRNA diagnostics relative to other CRISPR-based RNA detection tools. |
| first_indexed | 2024-03-11T09:40:25Z |
| format | Article |
| id | doaj.art-f48430cfb98944a782474642602a6f0c |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:40:25Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f48430cfb98944a782474642602a6f0c2023-11-16T17:03:24ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01243285710.3390/ijms24032857Harnessing the LdCsm RNA Detection Platform for Efficient microRNA DetectionZhenxiao Yu0Jianan Xu1Qunxin She2CRISPR and Archaea Biology Research Center, State Key Laboratory of Microbial Technology and Microbial Technology Institute, Shandong University, Qingdao 266237, ChinaCRISPR and Archaea Biology Research Center, State Key Laboratory of Microbial Technology and Microbial Technology Institute, Shandong University, Qingdao 266237, ChinaCRISPR and Archaea Biology Research Center, State Key Laboratory of Microbial Technology and Microbial Technology Institute, Shandong University, Qingdao 266237, ChinaIn cancer diagnosis, diverse microRNAs (miRNAs) are used as biomarkers for carcinogenesis of distinctive human cancers. Thus, the detection of these miRNAs and their quantification are very important in prevention of cancer diseases in human beings. However, efficient RNA detection often requires RT-PCR, which is very complex for miRNAs. Recently, the development of CRISPR-based nucleic acid detection tools has brought new promises to efficient miRNA detection. Three CRISPR systems can be explored for miRNA detection, including type III, V, and VI, among which type III (CRISPR-Cas10) systems have a unique property as they recognize RNA directly and cleave DNA collaterally. In particular, a unique type III-A Csm system encoded by <i>Lactobacillus delbrueckii</i> subsp. <i>bulgaricus</i> (LdCsm) exhibits robust target RNA-activated DNase activity, which makes it a promising candidate for developing efficient miRNA diagnostic tools. Herein, LdCsm was tested for RNA detection using fluorescence-quenched DNA reporters. We found that the system is capable of specific detection of miR-155, a microRNA implicated in the carcinogenesis of human breast cancer. The RNA detection system was then improved by various approaches including assay conditions and modification of the 5′-repeat tag of LdCsm crRNAs. Due to its robustness, the resulting LdCsm detection platform has the potential to be further developed as a better point-of-care miRNA diagnostics relative to other CRISPR-based RNA detection tools.https://www.mdpi.com/1422-0067/24/3/2857CRISPR-Cas10type III-AHD domainDNaseRNA detectionmiRNA |
| spellingShingle | Zhenxiao Yu Jianan Xu Qunxin She Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection International Journal of Molecular Sciences CRISPR-Cas10 type III-A HD domain DNase RNA detection miRNA |
| title | Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection |
| title_full | Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection |
| title_fullStr | Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection |
| title_full_unstemmed | Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection |
| title_short | Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection |
| title_sort | harnessing the ldcsm rna detection platform for efficient microrna detection |
| topic | CRISPR-Cas10 type III-A HD domain DNase RNA detection miRNA |
| url | https://www.mdpi.com/1422-0067/24/3/2857 |
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