한빛사논문
- 조회420
Donghyun Lim1,2,3, Vedagopuram Sreekanth1,2,3, Kurt J. Cox1,2,3, Benjamin K. Law1,2,3, Bridget K. Wagner1, Jeffrey M. Karp4,5,6,7 & Amit Choudhary1,2,3
1Chemical Biology and Therapeutics Science Program, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
2Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
3Divisions of Renal Medicine and Engineering, Brigham and Women’s Hospital, Boston, MA 02115, USA.
4Engineering in Medicine, Department of Medicine, Center for Regenerative Therapeutics, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA.
5Harvard−MIT Division of Health Sciences and Technology, MIT, Cambridge, MA 02139, USA.
6Proteomics Platform, Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
7Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA.
Corresponding Author: Amit Choudhary
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.
논문정보
- Research article
- 2020년 08월 (BRIC 등록일 2023-01-05)
- 국외 연구진
- 생명과학 > 생물공학
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