Methods and compositions for conferring cellular resistance to viral infection

A method of conferring viral resistance to a cell, comprising:

• delivering to the cell a first viral transfer ribonucleic acid (tRNA) variant, wherein the first viral tRNA variant binds to a first messenger ribonucleic acid (mRNA) codon and is charged with an amino acid not encoded by the first mRNA codon.

The method of any one of the preceding claims, wherein the first mRNA codon encodes an amino acid selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine.

The method of any one of the preceding claims, wherein the first amino acid is selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine, optionally wherein the first amino acid is leucine.

The method of any one of the preceding claims, wherein the second amino acid is selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine, optionally wherein the second amino acid is leucine.

The method of any one of the preceding claims, wherein the cell does not comprise an endogenous tRNA that binds to the first mRNA codon.

The method of any one of the preceding claims, wherein the cell expresses an essential gene encoding a protein that comprises a nonstandard amino acid.

The method of any one of the preceding claims, wherein the cell is a mammalian cell, a yeast cell, a plant cell, or a bacterial cell.

The method of any one of the preceding claims, wherein the cell further comprises an mRNA comprising the first mRNA codon to which the first viral tRNA variant binds.

The method of any one of the preceding claims, wherein the cell comprises an artificial genetic code in which an mRNA codon encodes an amino acid different from its natural amino acid identity.

The method of any one of the preceding claims, wherein the first and/or second viral tRNA variant comprises the sequence of any one of SEQ ID NO:

• 1-6 or is selected from the tRNA variants of Table 3.

A cell, comprising:

• an engineered nucleic acid encoding the first viral tRNA variant, wherein the first viral tRNA variant binds to a first messenger ribonucleic acid (mRNA) codon and is charged with an amino acid not encoded by the first mRNA codon, wherein the cell is resistant to viral infection.

An engineered cell, comprising:

• a first viral transfer ribonucleic acid (tRNA) variant, wherein the first viral tRNA variant binds to a first messenger ribonucleic acid (mRNA) codon and is charged with an amino acid not encoded by the first mRNA codon, wherein the cell is resistant to viral infection.

The cell of any one of the preceding claims, wherein the first mRNA codon encodes an amino acid selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine.

The cell of any one of the preceding claims, wherein the first amino acid is selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine, optionally wherein the first amino acid is leucine.

The cell of any one of the preceding claims, wherein the second amino acid is selected from alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, tryptophan, tyrosine, and valine, optionally wherein the second amino acid is leucine.

The cell of any one of the preceding claims, wherein the cell does not comprise an endogenous tRNA that binds to the first mRNA codon.

The cell of any one of the preceding claims, wherein the cell expresses an essential gene encoding a protein that comprises a nonstandard amino acid.

The cell of any one of the preceding claims, wherein the cell is a mammalian cell, a yeast cell, a plant cell, or a bacterial cell.

The cell of any one of the preceding claims, wherein the cell further comprises an mRNA comprising the first mRNA codon to which the first viral tRNA variant binds.

The cell of any one of the preceding claims comprising an artificial genetic code in which an mRNA codon encodes an amino acid different from its natural amino acid identity.

The cell of any one of the preceding claims, wherein the first and/or second viral tRNA variant comprises the sequence of any one of SEQ ID NOs:

• 1-6 or is selected from the tRNA variants of Table 3.