Modified nanopores, compositions comprising the same, and uses thereof

Abstract

Provided herein relate to modified or mutant forms of cytolysin A (ClyA) and compositions comprising the same. In particular, the modified or mutant forms of ClyA permits efficient capture and / or translocation of a target negative-charged molecule or polymer through the modified or mutant ClyA nanopores at low or physiological ionic strengths. Thus, methods for using the modified or mutant forms of ClyA and compositions, for example, for characterizing a target negatively-charged analyte, e.g., a target polynucleotide, are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 264,709 filed December 8, 2015 under 35 U.S.C. § 119(e), the contents of which are hereby incorporated by reference in their entirety. technical field [0003] Provided herein are modified or mutated forms of cytolysin A (ClyA) and compositions comprising the same. Also provided are methods of using said modified or mutated forms of ClyA and said compositions, for example, to characterize an analyte of interest, such as a polynucleotide of interest. Background technique [0004] Transmembrane pores such as nanopores have been used to recognize small molecules or folded proteins and to monitor chemical or enzymatic reactions at the single-molecule level. Electrophoretic translocation of DNA across nanopores reconstituted in artificial membranes holds great promise for practical applications such as DNA sequencing and biomarker identification. H...

AI Technical Summary

Problems solved by technology

However, translocation of double-stranded or single-stranded DNA through nanopores with negatively charged amino acids towards the inner surface is ineffective

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