Mutant pore

Patent metadata

Field	Value
Publication number	US-12371458-B2
Application number	US-202418440278-A
Country	US
Kind code	B2
Filing date	Feb 13, 2024
Priority date	Apr 6, 2016
Publication date	Jul 29, 2025
Grant date	Jul 29, 2025

Abstract

The invention relates to mutant forms of lysenin. The invention also relates to analyte characterisation using the mutant forms of lysenin.

First claim

The invention claimed is:

1. An apparatus comprising a transmembrane protein pore inserted into an in vitro membrane, wherein the transmembrane protein pore comprises at least one mutant lysenin monomer comprising a variant of the sequence set forth in SEQ ID NO: 2, wherein the variant comprises the S49K/R/L amino acid substitution.

2. The apparatus of claim 1 , wherein the transmembrane protein pore is a homo-oligomeric pore comprising the at least one mutant lysenin monomer.

3. The apparatus of claim 1 , wherein the transmembrane protein pore is a hetero-oligomeric pore comprising the at least one mutant lysenin monomer.

4. The apparatus of claim 1 , wherein the variant further comprises the E94 D/Q/G/A/K/R amino acid substitution.

5. An apparatus for characterizing a target analyte, the apparatus comprising a plurality of mutant lysenin monomers, wherein each mutant lysenin monomer comprises a variant of the sequence set forth in SEQ ID NO: 2, wherein the variant comprises the S49K/R/L amino acid substitution, and wherein the plurality of mutant lysenin monomers is inserted into a plurality of in vitro membranes.

6. The apparatus of claim 5 , wherein each of the plurality of in vitro membranes comprises a homo-oligomeric pore comprising the mutant lysenin monomers.

7. The apparatus of claim 5 , wherein each of the plurality of in vitro membranes comprises a hetero-oligomeric pore, and wherein at least one monomer of the hetero-oligomeric pore is the mutant lysenin monomer.

8. The apparatus of claim 5 , wherein each lysenin variant further comprises the E94 D/Q/G/A/K/R amino acid substitution.

9. A transmembrane protein pore comprising at least one mutant lysenin monomer, wherein the at least one mutant lysenin monomer comprises the amino acid sequence of SEQ ID NO: 2 having the S49K/R/L amino acid substitution.

10. The pore of claim 9 , wherein the pore is a hetero-oligomeric pore.

11. The pore of claim 9 , wherein the pore is a homo-oligomeric pore.

12. The pore of claim 9 , wherein the at least one mutant lysenin monomer further comprises the E94 D/Q/G/A/K/R amino acid substitution.

13. An apparatus produced by a method comprising: (i) obtaining a transmembrane protein pore comprising at least one mutant lysenin monomer, wherein the at least one mutant lysenin monomer comprises the amino acid sequence of SEQ ID NO: 2 having the S49K/R/L amino acid substitution and (ii) contacting the pore with an in vitro membrane such that the pore is inserted into the in vitro in vitro membrane.

14. The pore of claim 13 , wherein the at least one mutant lysenin monomer further comprises the E94 D/Q/G/A/K/R amino acid substitution.

Assignees

• Oxford Nanopore Tech Plc

Inventors

• Jayasinghe Lakmal Nishantha
• Bruce Mark John
• Mcneill Luke
• Nathani Ramiz Iqbal
• Singh Pratik Raj
• Wood Neil Roger
• Young Stephen Robert

Classifications

• C07K14/43504

  from invertebrates · CPC title

• A61K35/62

  Leeches; Worms, e.g. cestodes, tapeworms, nematodes, roundworms, earth worms, ascarids, filarias, hookworms, trichinella or taenia · CPC title

• C12Q2563/116

  electrical properties of nucleic acids, e.g. impedance, conductivity or resistance · CPC title

• G01N33/48721

  Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68) · CPC title

• G01N27/26

  by investigating electrochemical variables; by using electrolysis or electrophoresis · CPC title