Materials and methods for the synthesis of error-minimized nucleic acid molecules
US9771576B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Feb 1, 2013 |
| Grant date | Sep 26, 2017 |
| Priority date | — |
| Expiry date | Feb 1, 2033 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/21
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides materials and methods useful for error correction of nucleic acid molecules. In one embodiment of the invention, a first plurality of double-stranded nucleic acid molecules having a nucleotide mismatch are fragmented by exposure to a molecule having unidirectional mismatch endonuclease activity. The nucleic acid molecules are cut at the mismatch site or near the mismatch site, leaving a double-stranded nucleic acid molecule having a mismatch at the end or near end of the molecule. The nucleic acid molecule is then exposed to a molecule having unidirectional exonuclease activity to remove the mismatched nucleotide. The missing nucleotides can then be filled in by the action of, e.g., a molecule having DNA polymerase activity. The result is double-stranded nucleic acid molecules with a decreased frequency of nucleotide mismatches. Also provided are novel nucleic acid sequences encoding mismatch endonucleases, polypeptides encoded thereby, as well as nucleic acid constructs, transgenic cells, and various compositions thereof.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.