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Genetic code expansion in the mouse brain


Site-specific incorporation of non-natural amino acids into proteins, via genetic code expansion with pyrrolysyl tRNA synthetase (PylRS) and tRNAPylCUA pairs (and their evolved derivatives) from Methanosarcina sp., forms the basis of powerful approaches to probe and control protein function in cells and invertebrate organisms. Here we demonstrate that adeno-associated viral delivery of these pairs enables efficient genetic code expansion in primary neuronal culture, organotypic brain slices and the brains of live mice.

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Figure 1: AAV-mediated incorporation of non-natural amino acids into proteins in dissociated neurons and bioorthogonal labeling.
Figure 2: Incorporation of non-natural amino acids into proteins in SCN slices and live mice.


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This work was supported by the Medical Research Council (MRC), UK (MC_U105181009 and MC_UP_A024_1008, to J.W.C., MC_U105170643 to M.H.H.). V.B. is supported by an MRC case studentship (Nikon). We thank the MRC biomedical facility staff at ARES for their help.

Author information




J.W.C. defined the direction of research. R.J.E. designed the AAV vectors. V.B. and R.J.E. performed the experiments in rat cortical neurons. R.Z., E.S.M. and R.J.E. defined the amino acid delivery conditions in live mice. R.J.E. and T.P.K. performed the SCN slice experiments under the direction of M.H.H. R.Z. performed the labeling of brain sections containing 3 with 6 and performed microscopy. E.S.M. performed the experiments in live mice. T.S.E. provided amino acid 1. R.Z. provided amino acid 3, designed pharmacokinetic experiments, determined the plasma concentrations of 3 and analyzed pharmacokinetic data. N.P.B. performed quantitative microscopy and assisted imaging experiments. J.W.C. and R.J.E. wrote the paper with input from all authors.

Corresponding author

Correspondence to Jason W Chin.

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The authors declare no competing financial interests.

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Supplementary Results and Supplementary Figures 1–13. (PDF 4466 kb)

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Ernst, R., Krogager, T., Maywood, E. et al. Genetic code expansion in the mouse brain. Nat Chem Biol 12, 776–778 (2016).

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