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bioRxiv · 10.1101/2024.07.08.602493

Intracellular Protein Editing to Enable Incorporation of Non-Canonical Residues into Endogenous Proteins

Beyer, J.·Serebrenik, Y.·Toy, K.·Najar, M. A.·Raniszewski, N.·Shalem, O.·Burslem, G.
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Abstract

The ability to study proteins in a cellular context is crucial to our understanding of biology. Here, we report a new technology for "intracellular protein editing", drawing from intein- mediated protein splicing, genetic code expansion, and endogenous protein tagging. This protein editing approach enables us to rapidly and site specifically install residues and chemical handles into a protein of interest. We demonstrate the power of this protein editing platform to edit cellular proteins, inserting epitope peptides, protein-specific sequences, and non-canonical amino acids (ncAAs). Importantly, we employ an endogenous tagging approach to apply our protein editing technology to endogenous proteins with minimal perturbation. We anticipate that the protein editing technology presented here will be applied to a diverse set of problems, enabling novel experiments in live mammalian cells and therefore provide unique biological insights. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=85 SRC="FIGDIR/small/602493v1_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@8957b8org.highwire.dtl.DTLVardef@de02beorg.highwire.dtl.DTLVardef@1863cb4org.highwire.dtl.DTLVardef@1fb8bc0_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Beyer, J., Serebrenik, Y., Toy, K., Najar, M. A., Raniszewski, N., Shalem, O., Burslem, G.. 2024-07-08. Intracellular Protein Editing to Enable Incorporation of Non-Canonical Residues into Endogenous Proteins. https://doi.org/10.1101/2024.07.08.602493

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