Peptide fusion improves prime editing efficiency

Author(s)

Velimirovic, Minja ; Zanetti, Larissa C. ; Shen, Max W. ; Fife, James D. ; Lin, Lin ; Cha, Minsun ; Akinci, Ersin ; Barnum, Danielle ; Yu, Tian ; Sherwood, Richard I.

• Document type: Journal article
• Source: NATURE COMMUNICATIONS; v. 13, n. 1, p. 8-pg., 2022-06-18.

Abstract

Prime editing enables search-and-replace genome editing but is limited by low editing efficiency. Here the authors present PepSEq, a high-throughput method for screening a large library of peptides that influence prime editing efficiency. Prime editing enables search-and-replace genome editing but is limited by low editing efficiency. We present a high-throughput approach, the Peptide Self-Editing sequencing assay (PepSEq), to measure how fusion of 12,000 85-amino acid peptides influences prime editing efficiency. We show that peptide fusion can enhance prime editing, prime-enhancing peptides combine productively, and a top dual peptide-prime editor increases prime editing significantly in multiple cell lines across dozens of target sites. Top prime-enhancing peptides function by increasing translation efficiency and serve as broadly useful tools to improve prime editing efficiency. (AU)

• FAPESP's process: 17/25009-1 - Gene modulation of RIPK3 and MLKL and its impact in the sensibility to death by necroptosis
• Grantee: Larissa Cardoso Zanetti
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 19/13813-6 - High throughput mapping of MLKL transcriptional regulatory elements
• Grantee: Larissa Cardoso Zanetti
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate (Direct)