Compositions and methods of labeling mitochondrial nucleic acids and sequencing and analysis thereof

US12258615B2 · kind B2 · utility

Assignee

• King Abdullah University of Science and Technology · SA

Inventors

• Mo Li · Irvine, US
• Chongwei Bi · Thuwal, SA

Key dates

• Filing date: Aug 23, 2021
• Grant date: Mar 25, 2025
• Expiry date: Sep 24, 2042

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C40B40/08
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods of labeling, amplifying, and sequencing mitochondrial DNA are provided. The labeling methods typically include using polymerase-based extension of one or more unique molecular identifier (UMI) primers along a target mitochondrial DNA template, each UMI primer including a universal primer sequence, a unique molecular identifier sequence, and a mitochondrial DNA binding sequence. Amplification of the labeled-target mitochondrial DNA can include, for example, polymerase chain reaction (e.g., high-fidelity long-range). Methods of determining the sequence of labeled-target mitochondrial DNA can include use of a long-read sequencing platform. The methods may further including one or more of grouping sequences having the same UMI into one of more groups, determining the sequence of each labeled-target mitochondrial DNA by determining the consensus sequence of each group, and identifying polymorphisms in one or more of the labeled-target mitochondrial DNA. The methods can be used in, for example, preimplantation genetic testing.