A conjugation platform for CRISPR-Cas9 allows efficient β-cell engineering

作者: Donghyun Lim , Vedagopuram Sreekanth , Kurt J. Cox , Benjamin K. Law , Bridget K. Wagner

DOI: 10.1101/732354

关键词:

摘要: ABSTRACT Genetically fusing protein domains to Cas9 has yielded several transformative technologies; however, these fusions are polypeptidic, limited the termini and lack multivalent display, exclude diverse array of molecules. Here, we report a platform for site-specific display wide assortment molecules on both internal sites Cas9. Using this platform, endow with functionality effect precision genome edits, which involves efficient incorporation exogenously supplied single-stranded oligonucleotide donor (ssODN) at break site. We demonstrate that ssODN significantly increased edits over those bearing one or no ssODN, such is compatible large oligonucleotides rapid screening ssODNs. By hijacking insulin secretion machinery leveraging successfully engineer pancreatic β cells secrete protective immunomodulatory factor interleukin-10.

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