Patent ID: 9422550
Filing Date: 2016-08-23
CPC Classification: A61K,C12N,C40B

Claim Text:
1. A method for linking an mRNA molecule to a polypeptide, the method comprising: (a) providing an mRNA molecule comprising a nucleophilic nucleoside partner; (b) providing a translation system comprising a linking aminoacyl-tRNA comprising an amino acid residue linked to a linking tRNA by a covalent bond, wherein the linking tRNA comprises an anticodon comprising a crosslinker, wherein the crosslinker is an electrophilic nucleoside that reacts with a nucleophilic nucleoside partner via hybridization-triggered alkylation; (c) translating the mRNA molecule in the translation system to produce a polypeptide into which the amino acid residue, still linked to the linking tRNA, is incorporated; and (d) during or after step (c), crosslinking the crosslinker of the linking tRNA to the nucleophilic nucleoside partner of the mRNA molecule, thereby linking the mRNA molecule to the polypeptide through the linking tRNA, wherein the electrophilic nucleoside and the nucleophilic nucleoside partner are selected from the group consisting of: (i) 2-amino-6-vinylpurine and cytosine; (ii) 2-amino-6-vinylpurine and 4-thiouridine; (iii) 2-alpha-halomethyl adenosine and 2-thiouridine; (iv) 8-alpha-halomethyl purine and 4-thiouridine; (v) 2,8-alpha-halomethyl purine and 4-thiouridine; (vi) 5-methyl-N4,N4-ethanocytosine and cytosine; (vii) 2-amino-6-(1-ethylsulfinyl)vinyl purine nucleoside and cytosine; (viii) 4-amino-6-oxo-2-vinylpyrimidine nucleoside and uridine; and (ix) 4-amino-6-oxo-2-vinylpyrimidine-ethyl-C-nucleoside and uridine.