CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers
Abstract Recent advances in gene editing and precise regulation of gene expression based on CRISPR technologies have provided powerful tools for the understanding and manipulation of gene functions. Fusing RNA aptamers to the sgRNA of CRISPR can recruit cognate RNA-binding protein (RBP) effectors to...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-55957-0 |
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| author | Qiwen Su-Tobon Jiayi Fan Michael Goldstein Kevin Feeney Hongyuan Ren Patrick Autissier Peiyi Wang Yingzi Huang Udayan Mohanty Jia Niu |
| author_facet | Qiwen Su-Tobon Jiayi Fan Michael Goldstein Kevin Feeney Hongyuan Ren Patrick Autissier Peiyi Wang Yingzi Huang Udayan Mohanty Jia Niu |
| author_sort | Qiwen Su-Tobon |
| collection | DOAJ |
| description | Abstract Recent advances in gene editing and precise regulation of gene expression based on CRISPR technologies have provided powerful tools for the understanding and manipulation of gene functions. Fusing RNA aptamers to the sgRNA of CRISPR can recruit cognate RNA-binding protein (RBP) effectors to target genomic sites, and the expression of sgRNA containing different RNA aptamers permit simultaneous multiplexed and multifunctional gene regulations. Here, we report an intracellular directed evolution platform for RNA aptamers against intracellularly expressed RBPs. We optimize a bacterial CRISPR-hybrid system coupled with FACS, and identified high affinity RNA aptamers orthogonal to existing aptamer-RBP pairs. Application of orthogonal aptamer-RBP pairs in multiplexed CRISPR allows effective simultaneous transcriptional activation and repression of endogenous genes in mammalian cells. |
| format | Article |
| id | doaj-art-ef880aac3d984038ab25e87ac9c44063 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-ef880aac3d984038ab25e87ac9c440632025-01-12T12:31:23ZengNature PortfolioNature Communications2041-17232025-01-0116111310.1038/s41467-025-55957-0CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA AptamersQiwen Su-Tobon0Jiayi Fan1Michael Goldstein2Kevin Feeney3Hongyuan Ren4Patrick Autissier5Peiyi Wang6Yingzi Huang7Udayan Mohanty8Jia Niu9Department of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Biology, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeDepartment of Chemistry, Boston CollegeAbstract Recent advances in gene editing and precise regulation of gene expression based on CRISPR technologies have provided powerful tools for the understanding and manipulation of gene functions. Fusing RNA aptamers to the sgRNA of CRISPR can recruit cognate RNA-binding protein (RBP) effectors to target genomic sites, and the expression of sgRNA containing different RNA aptamers permit simultaneous multiplexed and multifunctional gene regulations. Here, we report an intracellular directed evolution platform for RNA aptamers against intracellularly expressed RBPs. We optimize a bacterial CRISPR-hybrid system coupled with FACS, and identified high affinity RNA aptamers orthogonal to existing aptamer-RBP pairs. Application of orthogonal aptamer-RBP pairs in multiplexed CRISPR allows effective simultaneous transcriptional activation and repression of endogenous genes in mammalian cells.https://doi.org/10.1038/s41467-025-55957-0 |
| spellingShingle | Qiwen Su-Tobon Jiayi Fan Michael Goldstein Kevin Feeney Hongyuan Ren Patrick Autissier Peiyi Wang Yingzi Huang Udayan Mohanty Jia Niu CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers Nature Communications |
| title | CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers |
| title_full | CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers |
| title_fullStr | CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers |
| title_full_unstemmed | CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers |
| title_short | CRISPR-Hybrid: A CRISPR-Mediated Intracellular Directed Evolution Platform for RNA Aptamers |
| title_sort | crispr hybrid a crispr mediated intracellular directed evolution platform for rna aptamers |
| url | https://doi.org/10.1038/s41467-025-55957-0 |
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