Jenna N Beyer, Kaitlyn Toy, George M Burslem. A Protein Editing Method to Modify Proteins in Live Mammalian Cells. Curr Protoc. 2026 Aug;6(8):e70443. doi: 10.1002/cpz1.70443. PMID: 42627710; PMCID: PMC13496354.

Abstract

Protein editing, as mediated by protein trans‐splicing, offers the ability to manipulate, label, and modify proteins in vitro and in live mammalian cells. This ability to observe and control proteins with this high resolution is a burgeoning area of interest within the chemical biology field. In this protocol, we outline the general approach for setting up a cellular protein editing experiment, emphasizing the planning process for a successful protein editing experiment. Additionally, we include methods to install and subsequently label a click chemistry handle p‐azido‐phenylalanine, a non‐canonical amino acid (ncAA) within the intein donor, such that this ncAA and any label can be edited into a protein in live mammalian cells. Overall, we anticipate that this protocol should serve as a generalizable roadmap for planning and carrying out a protein editing experiment.