Import of unnatural or modified nucleoside triphosphates into cells via nucleic acid triphosphate transporters
AssigneeThe Scripps Research Institute
InventorsFloyd E. Romesberg, Denis A. Malyshev
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Abstract
A recombinantly expressed nucleotide triphosphate transporter efficiently imports the triphosphates of unnatural nucleotides into cells, and the endogenous cellular machinery incorporates those nucleotides into cellular nucleic acids. UBPs can therefore form within the cell's nucleic acids. Moreover, neither the presence of the unnatural triphosphates nor the replication of the UBP represents a significant growth burden. The UBP is not efficiently excised by nucleic acid repair pathways, and therefore can be retained as long as the unnatural triphosphates are available in the growth medium. Thus, the resulting cell is the first organism to stably propagate an expanded genetic alphabet.
Claims40
An isolated prokaryotic cell comprising:
- (a) a heterologous nucleoside triphosphate transporter comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:
- 1;
- and
- (b) a first unnatural nucleotide comprising a base selected from:
- wherein the heterologous nucleoside triphosphate transporter is capable of transporting triphosphates of the first unnatural nucleotide into the cell.
An isolated prokaryotic cell comprising:
- (a) a heterologous nucleoside triphosphate transporter comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:
- 1;
- and
- (b) a first unnatural nucleotide,
- wherein the heterologous nucleoside triphosphate transporter is capable of transporting triphosphates of the first unnatural nucleotide into the cell.
An isolated bacterial cell comprising a heterologous nucleoside triphosphate transporter comprising an amino acid sequence that is at least about 85% identical to SEQ ID NO:
- 1, wherein the heterologous nucleoside triphosphate transporter is capable of transporting a triphosphate of an unnatural nucleotide into the isolated bacterial cell.
Dependencies are inferred from the claim text.