Yale University

Dieter Söll

Farren J Isaacs

Miriam Amiram

Michael Hohn

Caroline Aldag

Takahito Mukai

Adrian Haimovich

Non-naturally occurring tRNAand methods of using them for recombinant expression of proteins engineered to include one or more selenocysteine residues are disclosed. The non-naturally occurring tRNAcan be used for recombinant manufacture of selenocysteine containing polypeptides encoded by mRNA without the requirement of an SECIS element. In some embodiments, selenocysteine containing polypeptides are manufactured by co-expressing a non-naturally occurring tRNAa recombinant expression system, such as, with SerRS, EF-Tu, SelA, or PSTK and SepSecS, and an mRNA with at least one codon that recognizes the anticodon of the non-naturally occurring tRNA.

Compositions and methods for making selenocysteine containing polypeptides

Compositions, systems, and methods for preparation of polypeptides having multiple iterations of non-standard amino acids are provided. The compositions and method can be used to produce recombinant proteins at a greater yield than the same or similar polypeptides made using conventional compositions, systems, and methods. Accordingly, in some embodiments, the polypeptides are ones that could not be made using conventional methods and reagents, or could not be made a sufficient yield or purity to serve a practical purpose using conventional methods and reagents. Polypeptides made using the disclosed compositions, systems, and methods are also provided.

Compositions and methods of use thereof for making polypeptides with many instances of nonstandard amino acids

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Guilford, CT, United States of America

Dieter Söll