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US 12,571,017 B2Granted
Devices and methods for producing nucleic acids and proteins
AssigneeLife Technologies Corporation
InventorsClaudia Chiocchini, Axel Trefzer, Krishna Vattem, Phillip Kuhn
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Abstract
The present disclosure generally relates to devices, compositions and methods for designing and producing nucleic acid molecules and the production of encoded proteins using these nucleic acid molecules. In some aspect, the disclosure relates to automation for the in vitro generation of coding DNA molecules, the in vitro transcription of these DNA molecules to generate protein coding RNA molecules, and the in vitro translation of these protein coding RNA molecules to produce proteins.
Claims11
A method of producing a protein, the method comprising:
- a) designing a codon optimized nucleic acid molecule, wherein the codon optimized nucleic acid molecule encodes a mammalian protein, and wherein codons of the nucleic acid molecule are optimized for translation in a yeast,
- (b) generating oligonucleotides encoding subportions of the codon optimized nucleic acid molecule,
- (c) assembling the oligonucleotides generated in (b) to produce the codon optimized nucleic acid molecule,
- (d) contacting the codon optimized nucleic acid molecule assembled in (c) with a coupled in vitro translation/transcription reaction mixture under conditions that result in the production of (1) mRNA encoding the protein encoded by the codon optimized nucleic acid molecule and (2) the protein encoded by the codon optimized nucleic acid molecule,
- wherein the codons of the nucleic acid molecule are selected to avoid inverse complementary repeats that are capable of forming secondary structures in the mRNA produced in the coupled in vitro translation/transcription reaction mixture, and wherein the reaction mixture of (d) comprises mammalian cell components.
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Dependencies are inferred from the claim text.