Compositions and methods for detecting molecular targets on chromosomal DNA
US11492667B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 8, 2021 |
| Grant date | Nov 8, 2022 |
| Priority date | — |
| Expiry date | Jun 8, 2041 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6804
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Compositions, methods and kits are provided for identifying the presence and location of a target in chromosomal DNA. A nicking endonuclease fused to a binding domain that binds to a constant region of an antibody (NEFP) is provided that may be used for binding to a target directly or via an antibody that binds to the target. The target may be a protein or structural feature of the DNA and its presence and location may correspond to a phenotype and/or pathology in a biopsy or other cell sample for diagnostic purposes. The background is reduced by the addition of a glycoaminoglycan (GAG) that reversibly inhibits binding of the NEFP to DNA. Nick translation in the presence of a strand displacing polymerase enables the incorporation of tagged nucleotides that (i) blocks re-nicking; (ii) facilitates immobilization of DNA fragments around the target for sequencing; and/or (iii) enables dye labelling of the chromosomal DNA within the cell nuclei for analysis by microscopy.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.