Versatile 3′ Functionalization of CRISPR Single Guide RNA

Cody M. Palumbo, Jeton M. Gutierrez-Bujari, Henriette O'Geen, David J. Segal, Peter A Beal

Research output: Contribution to journalArticle

Abstract

Specific applications of CRISPR/Cas genome editing systems benefit from chemical modifications of the sgRNA. Herein we describe a versatile and efficient strategy for functionalization of the 3′-end of a sgRNA. An exemplary collection of six chemically modified sgRNAs was prepared containing crosslinkers, a fluorophore and biotin. Modification of the sgRNA 3′-end was broadly tolerated by Streptococcus pyogenes Cas9 in an in vitro DNA cleavage assay. The 3′-biotinylated sgRNA was used as an affinity reagent to identify IGF2BP1, YB1 and hnRNP K as sgRNA-binding proteins present in HEK293T cells. Overall, the modification strategy presented here has the potential to expand on current applications of CRISPR/Cas systems.

Original languageEnglish (US)
JournalChemBioChem
DOIs
StateAccepted/In press - Jan 1 2020

Keywords

  • click chemistry
  • nucleic acids
  • protein engineering
  • proteomics
  • RNA

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Versatile 3′ Functionalization of CRISPR Single Guide RNA'. Together they form a unique fingerprint.

  • Cite this

    Palumbo, C. M., Gutierrez-Bujari, J. M., O'Geen, H., Segal, D. J., & Beal, P. A. (Accepted/In press). Versatile 3′ Functionalization of CRISPR Single Guide RNA. ChemBioChem. https://doi.org/10.1002/cbic.201900736