Title:
METHOD FOR INTEGRATING LONG FOREIGN GENE INTO SAFE REGION OF HUMAN PLURIPOTENT STEM CELL AND ALLOWING SAME TO NORMALLY FUNCTION THEREIN
Document Type and Number:
WIPO Patent Application WO/2020/171222
Kind Code:
A1
Abstract:
The purpose of the present invention is to safely and efficiently introduce a cytotoxic gene into human pluripotent stem cell genome and then allow the gene to exert the function thereof to such an extent that undifferentiated cells, which remain after the differentiation into target cells, can be efficiently and exhaustively removed thereby. By homologous recombination using genome editing technology, a foreign (target) gene sequence was successfully and efficiently inserted into a safe harbor region of hPSC genome for the first time. Moreover, the following findings were obtained, namely: AAVS1 region that is defined as a safe harbor region is liable to show a lower gene expression level compared to random gene transfer using, for example, a lentiviral vector; in the homologous recombinant cells thus obtained, a promoter is required to have a strong expression ability; and a toxicity selective for undifferentiated cells can be achieved by employing, as a functional gene, a suicide gene correlating to cell growth activity and controlling the concentration of a prodrug corresponding thereto. As a result, the present invention successfully provides a mechanism whereby a gene is safely introduced into a safe harbor region of a genome by homologous recombination and, after inducing differentiation, undifferentiated cells alone can be exhaustively and selectively exterminated from the differentiated cells.
Inventors:
KOSAI KEN-ICHIRO (JP)
MITSUI KAORU (JP)
IDE KANAKO (JP)
MITSUI KAORU (JP)
IDE KANAKO (JP)
Application Number:
PCT/JP2020/007164
Publication Date:
August 27, 2020
Filing Date:
February 21, 2020
Export Citation:
Assignee:
UNIV KAGOSHIMA (JP)
International Classes:
C12N5/0735; C12N5/10; C12N15/09; C12N15/52; C12N15/54; C12N15/55; C12N15/85; C12Q1/02
Domestic Patent References:
| WO2015107888A1 | 2015-07-23 |
Foreign References:
| JP2018531612A | 2018-11-01 |
Other References:
IDE, KANAKO ET AL.: "Development of ''suicide gene insertion technology in genome editing' aiming to completely overcome tumorigenesis of pluripotent stem cells", LECTURE ABSTRACTS OF THE 18TH CONGRESS OF THE JAPANESE SOCIETY FOR REGENERATIVE MEDICINE, 22 February 2019 (2019-02-22)
Attorney, Agent or Firm:
NAGOYA INTERNATIONAL PATENT FIRM et al. (JP)
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