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- W3048776676 abstract "Abstract Genetically fusing protein domains to Cas9 has yielded several transformative technologies; however, the genetic modifications are limited to natural polypeptide chains at the Cas9 termini, which excludes a diverse array of molecules useful for gene editing. Here, we report chemical modifications that allow site-specific and multiple-site conjugation of a wide assortment of molecules on both the termini and internal sites of Cas9, creating a platform for endowing Cas9 with diverse functions. Using this platform, Cas9 can be modified to more precisely incorporate exogenously supplied single-stranded oligonucleotide donor (ssODN) at the DNA break site. We demonstrate that the multiple-site conjugation of ssODN to Cas9 significantly increases the efficiency of precision genome editing, and such a platform is compatible with ssODNs of diverse lengths. By leveraging the conjugation platform, we successfully engineer INS-1E, a β-cell line, to repurpose the insulin secretion machinery, which enables the glucose-dependent secretion of protective immunomodulatory factor interleukin-10." @default.
- W3048776676 created "2020-08-18" @default.
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- W3048776676 date "2020-08-13" @default.
- W3048776676 modified "2023-10-12" @default.
- W3048776676 title "Engineering designer beta cells with a CRISPR-Cas9 conjugation platform" @default.
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- W3048776676 doi "https://doi.org/10.1038/s41467-020-17725-0" @default.
- W3048776676 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7426819" @default.
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