Granted Patent
B2
US 12,275,761 · App. 17/194,821 · Granted Apr 15, 2025
Mutant CsgG pores
Inventors:
Stefan Howorka (London, GB); Han Remaut (Roosbeek, BE); Lakmal Jayasinghe (Oxford, GB); Elizabeth Jayne Wallace (Oxford, GB); James Clarke (Oxford, GB); Richard George Hambley (Oxford, GB); Jonathan Bankes Pugh (Oxford, GB)
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Abstract
The invention relates to mutant forms of CsgG. The invention also related to analyte detection and characterisation using CsgG.
Claims (7)
1. A method measuring a target analyte, comprising:
(a) contacting the target analyte with a transmembrane pore comprising at least one CsgG monomer such that the target analyte moves through the pore; and
(b) taking one or more electrical measurements as the target analyte moves through the pore.
2. The method of claim 1 , wherein the at least one CsgG monomer is a variant of SEQ ID NO: 390, said variant comprising: (a) mutations at one or more of the following positions N40, Q42, D43, E44, K49, Y51, S54, N55, F56, S57, Q62, E101, N102, E124, E131, R142, D149, T150, E185, R192, D195, E201 and E203; and/or (b) deletion of one or more of the following positions F48, K49, P50, Y51, P52, A53, S54, N55, F56 and S57.
3. The method of claim 2 , wherein the variant comprises any of the combinations of mutations/substitutions selected from the group consisting of: (i) one or more mutations at the following positions N40, D43, E44, S54, S57, Q62, R97, E101, E124, E131, R142, T150 and R192; (ii) mutations at Y51/N55, Y51/F56, N55/F56 or Y51/N55/F56; (iii) Q42R or Q42K; (iv) K49R; (v) N102R, N102F, N102Y or N102W; (vi) D149N, D149Q or D149R; (vii) E185N, E185Q or E185R; (viii) D195N, D195Q or D195R; (ix) E201N, E201Q or E201R; (x) E203N, E203Q or E203R; and (xi) deletion of one or more of the following positions F48, K49, P50, Y51, P52, A53, S54, N55, F56 and S57.
4. The method of claim 1 , wherein the target analyte is a target polynucleotide and step (a) further comprises contacting the polynucleotide with a polynucleotide binding protein such that the protein controls the movement of the polynucleotide through the pore.
5. The method of claim 4 , wherein the polynucleotide binding protein is a helicase or is derived from a helicase.
Assignments (4)
CHANGE OF NAME
Recorded Jan 14, 2022
From: OXFORD NANOPORE TECHNOLOGIES LIMITED
To: OXFORD NANOPORE TECHNOLOGIES PLC
Reel/Frame 058737/0664 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 25, 2021
From: HOWORKA, STEFAN
To: VIB VZW
Reel/Frame 056664/0032 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 25, 2021
From: JAYASINGHE, LAKMAL; WALLACE, ELIZABETH JAYNE; CLARKE, JAMES ANTHONY; PUGH, JONATHAN BANKES; HAMBLEY, RICHARD GEORGE
To: OXFORD NANOPORE TECHNOLOGIES LTD.
Reel/Frame 056664/0059 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Jun 25, 2021
From: REMAUT, HAN
To: VRIJE UNIVERSITEIT BRUSSEL
Reel/Frame 056664/0067 →
Priority Claims (3)
GB 1422079
· Dec 11, 2014
· national
GB 1506754
· Apr 21, 2015
· national
GB 1508287
· May 14, 2015
· national
Continuity (3)
Continuation
15507947
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