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Generation of programmable splicing factors using RNA-binding proteins that activate exon inclusion

January 3, 2024
in Science
Generation of programmable splicing factors using RNA-binding proteins that activate exon inclusion
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We developed luciferase-based splicing reporters to evaluate 718 RNA-binding proteins (RBPs) for the ability to activate exon inclusion. Our screen detected RBPs with no previously characterized roles in splicing and utilized RBP domains displaying potent exon activation to develop programmable splicing modulators with improved strength, generalizability and size over previous versions.

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Fig. 1: Reporter design and validation.

References

Gerstberger, S. et al. A census of human RNA-binding proteins. Nat. Rev. Genet. 15, 829–845 (2014). This review article describes the landscape of known human RNA-binding proteins at the time of publication.

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Queiroz, R. M. L. et al. Comprehensive identification of RNA–protein interactions in any organism using orthogonal organic phase separation (OOPS). Nat. Biotechnol. 37, 169–178 (2019). This paper reports a recent effort to comprehensively identify human RBPs.

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Du, M. et al. CRISPR artificial splicing factors. Nat. Commun. 11, 2973 (2020). This paper presents the Cas13-based CRISPR artificial splicing factor design on which we aimed to build.

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Chen, X. et al. Context-defined cancer co-dependency mapping identifies a functional interplay between PRC2 and MLL-MEN1 complex in lymphoma. Nat. Commun. 14, 4259 (2023). This paper presents a link between TRNAU1AP and PRPF39 in certain cancer cells.

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Bos, T. J. et al. Tethered function assays as tools to elucidate the molecular roles of RNA-binding proteins. In RNA Processing (ed. Yeo, G.) (2016). This chapter presents an overview of the tethered function assay approach and a discussion of its strengths and limitations.

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Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

This is a summary of: Schmok, J. C. et al. Large-scale evaluation of the ability of RNA-binding proteins to activate exon inclusion. Nat. Biotechnol. https://doi.org/10.1038/s41587-023-02014-0 (2024).

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Generation of programmable splicing factors using RNA-binding proteins that activate exon inclusion.
Nat Biotechnol (2024). https://doi.org/10.1038/s41587-023-02032-y

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Published: 02 January 2024

DOI: https://doi.org/10.1038/s41587-023-02032-y

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