Method for selecting a target nucleic acid sequence

US11352658B2 · kind B2 · utility

Assignee

• NAVINCI DIAGNOSTICS AB · SE

Inventors

• Ulf Landegren · Uppsala, SE
• Rachel Yuan Nong · Uppsala, SE

Key dates

• Filing date: Jul 31, 2015
• Grant date: Jun 7, 2022
• Expiry date: Jun 6, 2037

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6853
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention relates to a method for selecting a target region of interest (ROI) in a target nucleic acid molecule using a nucleic acid probe comprising a 3′ sequence capable of hybridising to a target nucleic acid molecule and acting as a primer for the production of a complement of the target ROI (i.e. by target templated extension of the primer), and a sequence capable of templating the circularisation and ligation of the extended probe comprising the reverse complement of the target ROI and a portion of the probe. The circularised molecule thus obtained contains the reverse complement of the target ROI and may be subjected to further analysis and/or amplification etc. The probe may be provided as an oligonucleotide comprising a stem-loop structure or as a partially double-stranded construct and comprises a single-stranded 3′ end region containing the target-binding site. A second binding site provided in the probe serves as the ligation template for circularisation, and the stem-loop structure, if present, is cleaved to render the second binding site available for hybridisation to the target complement. Also provided are probes and kits for carrying out such a method.