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Folly Beach Library
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Wando Mount Pleasant Library
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CRISPR-Cas9 cytidine and adenosine base editing of splice-sites mediates highly-efficient disruption of proteins in primary and immortalized cells.
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- Author(s): Kluesner, Mitchell G.; Lahr, Walker S.; Lonetree, Cara-lin; Smeester, Branden A.; Qiu, Xiaohong; Slipek, Nicholas J.; Claudio Vázquez, Patricia N.; Pitzen, Samuel P.; Pomeroy, Emily J.; Vignes, Madison J.; Lee, Samantha C.; Bingea, Samuel P.; Andrew, Aneesha A.; Webber, Beau R.; Moriarity, Branden S.
- Source:
Nature Communications; 4/23/2021, Vol. 12 Issue 1, p1-12, 12p- Subject Terms:
- Source:
- Additional Information
- Abstract: CRISPR-Cas9 cytidine and adenosine base editors (CBEs and ABEs) can disrupt genes without introducing double-stranded breaks by inactivating splice sites (BE-splice) or by introducing premature stop (pmSTOP) codons. However, no in-depth comparison of these methods or a modular tool for designing BE-splice sgRNAs exists. To address these needs, we develop SpliceR (http://z.umn.edu/spliceR) to design and rank BE-splice sgRNAs for any Ensembl annotated genome, and compared disruption approaches in T cells using a screen against the TCR-CD3 MHC Class I immune synapse. Among the targeted genes, we find that targeting splice-donors is the most reliable disruption method, followed by targeting splice-acceptors, and introducing pmSTOPs. Further, the CBE BE4 is more effective for disruption than the ABE ABE7.10, however this disparity is eliminated by employing ABE8e. Collectively, we demonstrate a robust method for gene disruption, accompanied by a modular design tool that is of use to basic and translational researchers alike. Base editors can inactivate splice sites or introduce stop codons into a gene sequence. Here the authors present SpliceR to design, rank, and test sgRNAs for efficient gene disruption in T cells. [ABSTRACT FROM AUTHOR]
- Abstract: Copyright of Nature Communications is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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