Please download the dossier by clicking on the dossier button x
×

SCARLESS DNA ASSEMBLY AND GENOME EDITING USING CRISPR/CPF1 AND DNA LIGASE

  • US 20190330659A1
  • Filed: 07/14/2017
  • Published: 10/31/2019
  • Est. Priority Date: 07/15/2016
  • Status: Abandoned Application
First Claim
Patent Images

1. A method for assembling gene constructs in vitro from a plurality of DNA fragments, said method comprising the steps of:

  • (a) providing a plurality of DNA fragments comprising a first and second DNA fragment, wherein said first DNA fragment comprises a sequence overlap of at least three nucleic acids anywhere within the second DNA fragment;

    (b) digesting the first DNA fragment with a Cpf1 CRISPR system, thereby creating a sticky DNA end at the 5′ and

    /or 3′

    of said first DNA fragment, wherein said digested first DNA fragment ceases to be a target for said Cpf1 CRISPR system;

    (c) annealing the sticky end of the digested first DNA fragment from step (b) to a second compatible sticky end on the second DNA fragment; and

    (d) ligating the annealed DNA fragments from step (c) together, resulting in a ligated product;

    wherein the resulting ligated product is an assembled construct;

    wherein steps (b) and (d) are conducted in the same reaction without needing to inactivate the Cpf1 CRISPR system.

View all claims
  • 3 Assignments
Timeline View
Assignment View
    ×
    ×