Mutant pore

Patent metadata

Field	Value
Publication number	US-12018326-B2
Application number	US-202117511391-A
Country	US
Kind code	B2
Filing date	Oct 26, 2021
Priority date	Mar 2, 2016
Publication date	Jun 25, 2024
Grant date	Jun 25, 2024

Abstract

The invention relates to mutant forms of CsgG. The invention also relates to analyte detection and characterization using CsgG.

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 CsgG monomer comprising a variant having 80% sequence identity to the amino acid sequence of SEQ ID NO: 2,wherein the variant comprises a substitution at position K94 selected from K94N, K94Q, K94R, K94F, K94Y, K94W, K94L, and K94S, and wherein the transmembrane pore facilitates translocation of an analyte through the transmembrane pore.

2. The apparatus of claim 1 , wherein the variant comprises K94Q or K94N.

3. The apparatus of claim 1 , wherein the variant further comprises; Q42K or Q42R; E44N or E44Q; L90R or L90K; N91R or N91K; I95R or I95K; A99R or A99K; E101H, E101K, E101N, E101Q or E101T; and/or Q114K.

4. The apparatus of claim 1 , wherein the variant further comprises
one or more of I41N, R93F/Y/W/L/I/V/N/Q/S, A98K/R, Q100K/R, G103F/W/S/N/K/R, T104R/K, A106R/K, I107R/K/W/F/Y/L/V, N108R/K, L113K/R, S115R/K, T117R/K, Y130W/F/H/Q/N, K135L/V/N/Q/S, E 170S/N/Q/K/R, S208V/I/F/W/Y/L/T, D233S/N/Q/K/R, D238S/N/Q/K/R and E244S/N/Q/K/R;
one or more mutations at the following positions R93, G103 and 1107 or one or more of R93F/Y/W/L/I/V/N/Q/S, G103F/W/S/N/K/R and I107R/K/W/F/Y/L/V;
one or more mutations at the following positions I41, T104, A106, N108, L113, S115, T117, E170, D233, D238 and E244 or one or more of I41N, T104R/K, A106R/K, N108R/K, L113K/R, S115R/K, T117R/K, E170S/N/Q/K/R, D233S/N/Q/K/R, D238S/N/Q/K/R and E244S/N/Q/K/R;
one or more mutations at the following positions (i) A98, (ii) Q100, (iii) G103 and (iv) 1107 or one or more of (i) A98R/K, (ii) Q100K/R, (iii) G103K/R and (iv) I107R/K;
one or more mutations at the following positions Y130, K135 and S208 or one or more of Y130W/F/H/Q/N, K135L/V/N/Q/S and R142Q/S;
one or more one or more of (i) Q87N/R/K, (ii) K94R/F/Y/W/L/S/N, (iii) R97F/Y/W/V/I/K/S/Q/H, (iv) N102K/Q/L/I/V/S/H and (v) R110F/G/N;
one or more of (i) D43S, (ii) E44S, (iii) N91K/R, (iv) Q114R/K and (v) D248S/N/Q/K/R;
one or more of Q87R/K, E101I/L/A/H and N102K;
F48S/N/Q/Y/W/I/V; or
F48H/R/K.

5. The apparatus of claim 1 , wherein the variant further comprises R192D/Q/F/S/T/N/E or R192D/Q/F/S/T.

6. The apparatus of claim 1 , wherein the variant further comprises: R97W; R93W; R93Y and R97Y; F191T; or deletion of V105, A106 and I107.

Assignees

• Oxford Nanopore Tech Plc

Inventors

• Jayasinghe Lakmal
• Wallace Elizabeth Jayne

Classifications

• 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

• C12Q2565/631

  being a biochannel or pore · CPC title

• C07K14/245 Primary

  Escherichia (G) · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title