The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.

The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.

Date of Patent:
May. 03, 2022

Filed:

May. 25, 2019
Applicant:

Shanghai Ocean University, Shanghai, CN;

Inventors:

Qinghua Zhang, Shanghai, CN;

Ce Ji, Shanghai, CN;

Assignee:
Attorney:
Primary Examiner:
Int. Cl.
CPC ...
C12N 15/00 (2006.01); A01K 67/027 (2006.01); C07K 14/48 (2006.01); C12N 15/873 (2010.01); C12N 15/90 (2006.01); A61K 38/17 (2006.01);
U.S. Cl.
CPC ...
A01K 67/0276 (2013.01); C07K 14/48 (2013.01); C12N 15/873 (2013.01); C12N 15/902 (2013.01); A01K 2207/15 (2013.01); A01K 2217/05 (2013.01); A01K 2217/075 (2013.01); A01K 2227/40 (2013.01); A61K 38/1706 (2013.01); C12N 2310/20 (2017.05);
Abstract

A method of constructing a zebrafish notch1a mutant using CRISPR/Cas9 technique. The method includes: determining a target for knocking out notch1a; using primers T7-notch1a-sfd and tracr rev for PCR amplification with a pUC19-gRNA scaffold plasmid as a template; transcribing PCR product in vitro followed by purification to obtain gRNA; and microinjecting the gRNA and a Cas9 mRNA into a zebrafish embryo followed by culture to obtain an notch1a mutant of stable inheritance. The invention selects a specific target and utilizes CRISPR/Cas9 technique to knock out the notch1a in the zebrafish without destroying other genes, generating the zebrafish notch1a mutant. Moreover, the invention also discloses the phenotype of the zebrafish notch1a mutant, which plays a significant role in studying the effect of the Notch1a receptor in the Notch signaling pathway.


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