Title: Compositions and methods for making selenocysteine containing polypeptides

Abstract

Non-naturally occurring tRNA.sup.Sec and methods of using them for recombinant expression of proteins engineered to include one or more selenocysteine residues are disclosed. The non-naturally occurring tRNA.sup.Sec can 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 tRNA.sup.Sec a recombinant expression system, such as E. coli, 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.sup.Sec.

Inventors:

Soll, Dieter; Aldag, Caroline; Hohn, Michael

Issue Date:

2016-10-11

Research Org.:

Yale Univ., New Haven, CT (United States)

Sponsoring Org.:

USDOE

OSTI Identifier:

1328702

Patent Number(s):

9464288

Application Number:

14/131,382

Assignee:

Yale University (New Haven, CT)

Patent Classifications (CPCs):

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

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C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N9/14 - Hydrolases
C12N9/10 - Transferases
C12N15/113 - Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides
C12N9/1007 - {Methyltransferases

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

C12P21/00 - Preparation of peptides or proteins

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N2310/531 - Stem-loop

C - CHEMISTRY C07 - ORGANIC CHEMISTRY C07K - PEPTIDES

C07K14/47 - from mammals

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N9/0004 - {Oxidoreductases

C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES

C12Y209/01001 - L-Seryl-tRNA

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

C12N15/11 - DNA or RNA fragments
C12N9/93 - {Ligases
C12N9/0008 - {acting on the aldehyde or oxo group of donors

C - CHEMISTRY C12 - BIOCHEMISTRY C12Y - ENZYMES

C12Y306/05003 - Protein-synthesizing GTPase
C12Y601/01011 - Serine--tRNA ligase

C - CHEMISTRY C12 - BIOCHEMISTRY C12P - FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {

C12P21/02 - having a known sequence of two or more amino acids, e.g. glutathione

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DOE Contract Number:  

FG02-98ER20311

Resource Type:

Patent

Resource Relation:

Patent File Date: 2012 Jul 11

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY