Human embryonic stem cell line capable of controlling gene expression and application thereof

A technology for embryonic stem cells and gene regulation, applied to cells modified by introducing foreign genetic material, genetic engineering, using vectors to introduce foreign genetic material, etc., to achieve high-efficiency gene knock-in and reduce difficulty
CN108559731AInactive Publication Date: 2018-09-21SOUTHERN MEDICAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN ยท China
Patent Type
Applications(China)
Current Assignee / Owner
SOUTHERN MEDICAL UNIVERSITY
Publication Date
2018-09-21
Estimated Expiration
Not applicable ยท inactive patent

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Abstract

The invention discloses a human embryonic stem cell line capable of controlling gene expression and application thereof. The human embryonic stem cell line capable of controlling gene expression comprises a controlled expression exogenous fusion protein system and at least one guide RNA, and the controlled expression exogenous fusion protein system comprises a tetracycline inducible expression system and fusion protein formed by Cas9 protein and p300 core domain with transcriptional activation activity; each guide RNA is a 14bp short data sequence of a targeting gene promoter region or a 20bplong data sequence of a genome region. Accordingly, expression of Cas9-p300 in human embryonic stem cells can be controlled in real time, the specific gene activation and cutting double functions of human embryonic stem cell differentiation in different stages are achieved, and application of the human embryonic stem cells in the fields of medicine science and lift science is greatly extended.
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Description

technical field

[0001] The invention belongs to the field of genetic engineering and biotechnology, and relates to a Cas9-p300 human embryonic stem cell line with a controllable activation and cutting dual system mediated by CRISPR / Cas9, a construction method and application thereof. Background technique

[0002] Human embryonic stem cells are pluripotent cells, derived from the inner cell mass of early embryos (4-5d), with the potential of self-renewal and multilineage differentiation. In 1998, American scientist James Thomson successfully cultured human embryonic stem cells in vitro for the first time, and the scientific research and clinical application of human embryonic stem cells also began. In current research, embryonic stem cells have successfully differentiated into trophoblast cells, nerve cells, liver cells, chondrocytes, hematopoietic cells, cardiomyocytes and other cells, and have also successfully established individual development models. Clinical treatments...

Claims

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