Alpha-hemolysin variants and uses thereof

US10351908B2 · kind B2 · utility

Assignee

• Roche Sequencing Solutions, Inc. · US

Inventors

• Timothy Craig · Campbell, US
• Mark Ambroso · Escondido, US
• Corissa Harris · Mountain View, US
• Matthew DiPietro · Gilroy, US
• Yaozhong Zou · Sunnyvale, US
• Marshall Porter · Santa Clara, US

Key dates

• Filing date: Apr 20, 2017
• Grant date: Jul 16, 2019
• Expiry date: Apr 20, 2037

Classification

• Technology area (CPC G): Physics
• CPC primary: G01N2333/31
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

Described herein are variants of alpha-hemolysin having at least one mutation, such as a mutation to a positive charge. In certain examples, the mutation is selected from V149K, E287R, H35G, T109K, P151K, K147N, E111N, M113A, or combinations thereof in the mature, wild-type alpha-hemolysin amino acid sequence. The α-hemolysin variants may also include a substitution at H144A and/or a series of glycine residues spanning residues 127 to 131 of the mature, wild-type alpha hemolysin. Also provided are nanopore assemblies including the alpha-hemolysin variants, the assembly having a decreased time-to-thread. The decreased time-to-thread, for example, increases DNA sequencing efficiency and accuracy.