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. Author manuscript; available in PMC: 2020 Feb 18.
Published in final edited form as: ACS Chem Biol. 2018 Sep 5;13(9):2375–2386. doi: 10.1021/acschembio.8b00520

Figure 5.

Figure 5.

Expanding the genetic code in Mus musculus. (A) Diagram of UAA incorporation into proteins in mice via AAV transduction of the expression cassette or generation of stable lines through genome integration, with the UAA introduced either intracranially or through the drinking water. (B) Chemical structure of PropK and AcK. (C) Fluorescence imaging showing incorporation of PropK into sfGFP151TAG in coronal sections of the hypothalamus and suprachiasmatic nucleus. Scale bar: 500 μm.61 (D) DNA constructs used for generation of a transgenic mouse line for UAA incorporation. (E) Western blotting analysis of anti-FLAG-immunoprecipitated proteins from AcK-encoding sfGFP151TAG mice, demonstrating successful genetic code expansion.80 (F) Fluorescence imaging showing incorporation of AcK into GFP39TAG in skeletal muscle, liver, and lung tissues of live mice. Scale bar: 200 μm. Adapted with permission from Nat. Chem. Biol. 2016, 12, 776 and Nat. Commun. 2017, 8, 14568. Copyright 2016 and 2017 Nature Publishing Group.