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:
Nov. 22, 2022

Filed:

Apr. 11, 2017
Applicant:

Applied Stemcell, Inc., Milpitas, CA (US);

Inventors:

Ling-jie Kong, Foster City, CA (US);

Ruby Yanru Tsai, San Jose, CA (US);

Assignee:

APPLIED STEMCELL, INC., Milpitas, CA (US);

Attorneys:
Primary Examiner:
Int. Cl.
CPC ...
C12N 15/10 (2006.01); C12N 15/09 (2006.01); C12N 15/63 (2006.01); C12N 15/64 (2006.01); C12N 15/90 (2006.01); C07H 21/04 (2006.01); C12N 9/12 (2006.01); C12N 15/11 (2006.01); C12N 15/66 (2006.01);
U.S. Cl.
CPC ...
C12N 15/10 (2013.01); C07H 21/04 (2013.01); C12N 9/1211 (2013.01); C12N 15/09 (2013.01); C12N 15/102 (2013.01); C12N 15/11 (2013.01); C12N 15/63 (2013.01); C12N 15/64 (2013.01); C12N 15/66 (2013.01); C12N 15/90 (2013.01); C12N 15/907 (2013.01); C12Y 207/01021 (2013.01);
Abstract

Provided is a method for knocking in a gene of interest to a cell. The genome of the cell contains a negative selectable marker, e.g., a thymidine kinase gene flanked by a pair of recombinase recognition sites (RRS), e.g., attP. The method involves introducing into the cell a targeting construct that contains a gene of interest flanked by a second pair of RRS, e.g., attB. The targeting construct also contains in the vector backbone a negative selectable marker, e.g., thymidine kinase gene. When a recombinase recognizing the RRS is expressed, the recombination events between the two pairs of RRS result in the site-specific integration of the gene of interest in the genome of the cell. Upon selection based on the negative selectable marker, the parental cells, cells with undesired integration, e.g., random integration, or the integration of the vector backbone are removed.


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