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US 10,883,109 B2Granted
Methods of modulating mRNA stability and protein expression
AssigneeCase Western Reserve University
InventorsJeffery M. Coller, Kristian E. Baker
Life of this patent
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Abstract
A synthetic cDNA which encodes a protein wherein at least one optimal or non-optimal codon in a wild type DNA encoding the protein has been replaced respectively with one or more non-optimal codons or optimal codons encoding the same amino acid.
Claims6
A method of modulating the expression of a recombinant protein in a host cell, the method comprising:
- identifying optimal and non-optimal codons in a wild type cDNA sequence that encodes the protein,
- preparing a synthetic cDNA sequence by replacing one or more of the optimal codons of the wild type cDNA with a non-optimal codon encoding the same amino add as the replaced optimal codon or replacing one or more of the non-optimal codons of the wild type cDNA with an optimal codon encoding the same amino add as the replaced non-optimal codon,
- transfecting the host cell with the synthetic cDNA, wherein the replacement of the one or more codons from the cDNA sequence enhances expression of the recombinant protein in the host cell at least about 10% compared to the nucleic sequence prior to replacement, wherein the one or more optimal codons are selected from the group consisting of pct (Alanine), ggt (Glycine), gtc (Valine), ttg (Leucine), gtt (Valine), gcc (Alanine), cca (Proline), act (Threonine), tct (Serine), tcc (Serine), acc (Threonine), atc (Isoleucine), aag (Lysine), tac (Tyrosine), ttc (Phenylalanine), gaa (Glutamic Acid), cgt (Arginine), caa (Glutamine), cac (Histidine), aac (Asparagine), gac (Aspartic Add), att (Isoleucine), aga (Arginine), and tgt (Cysteine);
- and the one or more non-optimal codons are selected from the group consisting of cct (Proline), ggc (Glycine), igg (Tryptophan), tta (Leucine), gat (Aspartic Add), atg (Methionine), ttt (Phenylalanine), tgc (Cysteine), cat (Histidine), gca (Alanine), tat (Tyrosine), ccc (Proline), ggg (Glycine), gtg (Valine), gcg (Alanine), cgc (Arginine), tca (Serine), gag (Glutamic Acid), gga (Glycine), tcg (Serine), cgg (Arginine), aat (Asparagine), ctt (Leucine), cta (Leucine), cag (Glutamine), ctc (Leucine), aca (Threonine), age:
- (Serine), aaa (Lysine), agt (Serine), acg (Threonine), ctg (Leucine), ccg (Proline), gta (Valine), agg (Arginine), cga (Arginine), and ata (isoleucine), and
- expressing the recombinant protein.
3 more elements
Dependencies are inferred from the claim text.