Macromolecule delivery to nanowells

• US 10,550,423 B2
• Filed: 10/30/2017
• Issued: 02/04/2020
• Est. Priority Date: 12/29/2011
• Status: Active Grant

• Current Assignee: Ibis Biosciences Incorporated
• Original Assignee: Ibis Biosciences Incorporated
• Inventors: Gray, Phillip N.
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US15/797,982
• Publication Number:
  US 20180119205A1
• Time in Patent Office: 827 Days
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2521/543 Immobilised enzyme(s)
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins
• CPC Class Codes: C12Q 2565/631 being a biochannel or pore

Abstract

Provided herein is technology relating to depositing and/or placing a macromolecule at a desired site for an assay and particularly, but not exclusively, to methods and systems for transporting a macromolecule such as a protein, a nucleic acid, or a protein:nucleic acid complex to an assay site, such as the bottom of a nanopore, a nanowell, or a zero mode waveguide.

Claims

1. A system for transporting a macromolecule, the system comprising:

a) a transport guide comprising an actin filament or a microtubule with one end attached to a bottom of a zero mode wave guide well or nanowell;

b) a molecular motor for transporting the macromolecule along the transport guide wherein said molecular motor is a myosin, kinesin, or dynein; and

c) a linking domain linking the macromolecule to the molecular motor wherein said macromolecule is a DNA, a DNA polymerase or a DNA polymerase/DNA complex attached to said molecular motor, said molecular motor is configured to transport said DNA, said DNA polymerase or said DNA polymerase/DNA complex to said bottom of a zero mode wave guide well or nanowell.

2. The system of claim 1 further comprising a phospholinked nucleotide at said zero mode waveguide or nanowell.

3. The system of claim 1 further comprising an anchor to maintain the macromolecule at said zero mode wave guide well or nanowell.

4. The system of claim 1 wherein said linking domain is selected from the group consisting of a myosin binding domain and a microtubule associated protein binding domain.