FUSION PROTEIN PRODUCING POINT MUTATION IN CELL, AND PREPARATION AND USE THEREOF

Title:

FUSION PROTEIN PRODUCING POINT MUTATION IN CELL, AND PREPARATION AND USE THEREOF

Document Type and Number:

WIPO Patent Application WO/2017/215619

Kind Code:

A1

Abstract:

Provided are a fusion protein producing a point mutation in a cell, and preparation and use thereof. The fusion protein includes or is formed by a cytosine deaminase and Cas enzyme in which the nuclease activity is deficient but the helicase activity retains. Also provided are the coding sequence of the fusion protein, a polynucleotide sequence containing the coding sequence, a nucleic acid construct containing the polynucleotide sequence, the corresponding host cell, a method for producing the point mutation in the cell, and a kit, etc. The method can achieve site-directed mutagenesis, and at the same time, achieve high mutation efficiency and multiple mutation combinations in a particular gene region.

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Inventors:

CHANG XING (CN)

Application Number:

PCT/CN2017/088369

Publication Date:

December 21, 2017

Filing Date:

June 15, 2017

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Assignee:

SHANGHAI INST BIOLOGICAL SCIENCES CAS (CN)

International Classes:

C12N9/78; C07K19/00; C12N15/09

Domestic Patent References:

WO2015089406A1	2015-06-18
WO2015133554A1	2015-09-11
WO2016022363A2	2016-02-11

Foreign References:

CN105518146A

2016-04-20

Other References:

MA, Y.Q. ET AL.: "Targeted AID-Mediated Mutagenesis (TAM) Enables Efficient Genomic Diversification in Mammalian Cells", NATURE METHODS, vol. 13, no. 12, 10 October 2016 (2016-10-10), pages 1029 - 1035, XP002778319
LEE, C.M. ET AL.: "The Neisseria Meningitidis CRISPR-Cas9 System Enables Specific Genome Editing in Mammalian Cells", MOLECULAR THERAPY, vol. 24, no. 3, 16 February 2016 (2016-02-16), pages 645 - 654, XP055449590

Attorney, Agent or Firm:

SHANGHAI PATENT & TRADEMARK LAW OFFICE, LLC (CN)

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