Methods for generating libraries with co-varying regions of polynuleotides for genome modification
US10287590B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Dec 24, 2014 |
| Grant date | May 14, 2019 |
| Priority date | — |
| Expiry date | Dec 24, 2034 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/64
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The invention provides methods for introducing co-varying paired nucleic acids into a vector such under separate transcriptional control. An oligonucleotide is synthesized including the paired nucleic acids. The oligonucleotide is then assembled with a spacer nucleic acid encoding a promoter. After assembly the spacer nucleic acid is to one side of the oligonucleotide encoding the paired segments. However, on circularization of the assembled nucleic acid and cleavage between the DNA segments the spacer oligonucleotide and its components now occur between the paired segments. The resulting nucleic acid can now be cloned into a vector with a single step, such that each of the paired nucleic acid segments is linked to its own promoter. The present method can readily be extended to library screening without proportionately increasing the effort.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.