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A dual-constriction biological nanopore resolves homonucleotide sequences with high fidelity.

Abstract：

:Single-molecule long-read DNA sequencing with biological nanopores is fast and high-throughput but suffers reduced accuracy in homonucleotide stretches. We now combine the CsgG nanopore with the 35-residue N-terminal region of its extracellular interaction partner CsgF to produce a dual-constriction pore with improved signal and base-calling accuracy for homopolymer regions. The electron cryo-microscopy structure of CsgG in complex with full-length CsgF shows that the 33 N-terminal residues of CsgF bind inside the β-barrel of the pore, forming a defined second constriction. In complexes of CsgG bound to a 35-residue CsgF constriction peptide, the second constriction is separated from the primary constriction by ~25 Å. We find that both constrictions contribute to electrical signal modulation during single-stranded DNA translocation. DNA sequencing using a prototype CsgG-CsgF protein pore with two constrictions improved single-read accuracy by 25 to 70% in homopolymers up to 9 nucleotides long.

journal_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Van der Verren SE,Van Gerven N,Jonckheere W,Hambley R,Singh P,Kilgour J,Jordan M,Wallace EJ,Jayasinghe L,Remaut H

doi

10.1038/s41587-020-0570-8

subject

Has Abstract

pub_date

2020-12-01 00:00:00

pages

1415-1420

issue

12

eissn

1087-0156

issn

1546-1696

pii

10.1038/s41587-020-0570-8

journal_volume

38

pub_type

杂志文章

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