Engineering designer beta cells with a CRISPR-Cas9 conjugation platform

Donghyun Lim, Vedagopuram Sreekanth, Kurt J. Cox, Benjamin K. Law, Bridget K. Wagner, Jeffrey M. Karp and Amit Choudhary ()
Additional contact information
Donghyun Lim: Broad Institute of MIT and Harvard
Vedagopuram Sreekanth: Broad Institute of MIT and Harvard
Kurt J. Cox: Broad Institute of MIT and Harvard
Benjamin K. Law: Broad Institute of MIT and Harvard
Bridget K. Wagner: Broad Institute of MIT and Harvard
Jeffrey M. Karp: Center for Regenerative Therapeutics, Brigham and Women’s Hospital, Harvard Medical School
Amit Choudhary: Broad Institute of MIT and Harvard

Nature Communications, 2020, vol. 11, issue 1, 1-11

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.

Date: 2020
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41467-020-17725-0 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-17725-0

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-17725-0

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().