Optimization of in vitro isolation of nucleic acids using site-specific nucleases
US12305218B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Nov 18, 2019 |
| Grant date | May 20, 2025 |
| Priority date | — |
| Expiry date | Oct 22, 2042 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N9/22
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention relates to a method of preparing a nucleic acid molecule comprising a target region for isolation from a sample, said method comprising the steps of contacting a population of nucleic acid molecules with a first Class 2 Type V Cas protein-gRNA complex, wherein the gRNA comprises a guide segment that is complementary to a first sequence within a nucleic acid molecule, said first sequence being located adjacent to said target region, thereby forming a Class 2 Type V Cas protein-gRNA-nucleic acid complex, and a protector molecule, contacting the population of nucleic acid molecules with at least one enzyme having exonuclease activity, recovering said nucleic acid molecule comprising the target region from the Class 2 Type V Cas protein-gRNA-nucleic acid complex formed in step a), and contacting the nucleic acid molecule of step c) with a processive polymerase, preferably a mesophilic or thermophilic polymerase, even more preferably a T4 DNA polymerase.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.