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Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers
The development of RNA imaging strategies in live cells is essential to improve our understanding of their role in various cellular functions. We report an efficient RNA imaging method based on the CRISPR-dPspCas13b system with fluorescent RNA aptamers in sgRNA (CasFAS) in live cells. Using modified...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728512/ https://www.ncbi.nlm.nih.gov/pubmed/36540819 http://dx.doi.org/10.1039/d2sc04656c |
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author | Tang, Heng Peng, Junran Peng, Shuang Wang, Qi Jiang, Xin Xue, Xiaocheng Tao, Yanxin Xiang, Limin Ji, Quanjiang Liu, Song-Mei Weng, Xiaocheng Zhou, Xiang |
author_facet | Tang, Heng Peng, Junran Peng, Shuang Wang, Qi Jiang, Xin Xue, Xiaocheng Tao, Yanxin Xiang, Limin Ji, Quanjiang Liu, Song-Mei Weng, Xiaocheng Zhou, Xiang |
author_sort | Tang, Heng |
collection | PubMed |
description | The development of RNA imaging strategies in live cells is essential to improve our understanding of their role in various cellular functions. We report an efficient RNA imaging method based on the CRISPR-dPspCas13b system with fluorescent RNA aptamers in sgRNA (CasFAS) in live cells. Using modified sgRNA attached to fluorescent RNA aptamers that showed reduced background fluorescence, this approach provides a simple, sensitive way to image and track endogenous RNA with high accuracy and efficiency. In addition, color switching can be easily achieved by changing the fluorogenic dye analogues in living cells through user-friendly washing and restaining operations. CasFAS is compatible with orthogonal fluorescent aptamers, such as Broccoli and Pepper, enabling multiple colors RNA labeling or intracellular RNA–RNA interaction imaging. Finally, the visualization of severe fever with thrombocytopenia syndrome virus (SFTSV) was achieved by CasFAS, which may facilitate further studies on this virus. |
format | Online Article Text |
id | pubmed-9728512 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-97285122022-12-19 Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers Tang, Heng Peng, Junran Peng, Shuang Wang, Qi Jiang, Xin Xue, Xiaocheng Tao, Yanxin Xiang, Limin Ji, Quanjiang Liu, Song-Mei Weng, Xiaocheng Zhou, Xiang Chem Sci Chemistry The development of RNA imaging strategies in live cells is essential to improve our understanding of their role in various cellular functions. We report an efficient RNA imaging method based on the CRISPR-dPspCas13b system with fluorescent RNA aptamers in sgRNA (CasFAS) in live cells. Using modified sgRNA attached to fluorescent RNA aptamers that showed reduced background fluorescence, this approach provides a simple, sensitive way to image and track endogenous RNA with high accuracy and efficiency. In addition, color switching can be easily achieved by changing the fluorogenic dye analogues in living cells through user-friendly washing and restaining operations. CasFAS is compatible with orthogonal fluorescent aptamers, such as Broccoli and Pepper, enabling multiple colors RNA labeling or intracellular RNA–RNA interaction imaging. Finally, the visualization of severe fever with thrombocytopenia syndrome virus (SFTSV) was achieved by CasFAS, which may facilitate further studies on this virus. The Royal Society of Chemistry 2022-11-08 /pmc/articles/PMC9728512/ /pubmed/36540819 http://dx.doi.org/10.1039/d2sc04656c Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by-nc/3.0/ |
spellingShingle | Chemistry Tang, Heng Peng, Junran Peng, Shuang Wang, Qi Jiang, Xin Xue, Xiaocheng Tao, Yanxin Xiang, Limin Ji, Quanjiang Liu, Song-Mei Weng, Xiaocheng Zhou, Xiang Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title | Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title_full | Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title_fullStr | Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title_full_unstemmed | Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title_short | Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers |
title_sort | live-cell rna imaging using the crispr-dcas13 system with modified sgrnas appended with fluorescent rna aptamers |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9728512/ https://www.ncbi.nlm.nih.gov/pubmed/36540819 http://dx.doi.org/10.1039/d2sc04656c |
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