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Channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and construction method thereof

A neural stem cell and genetic engineering technology, which is applied in the field of ChR2-GFP genetically engineered neural stem cell lines and their construction, to achieve the effect of improving experimental efficiency

Inactive Publication Date: 2013-03-20
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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Problems solved by technology

Various studies currently provide only indirect evidence in terms of morphology and have not yet clarified the issue functionally, especially at the level of synapses that transmit information

Method used

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  • Channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and construction method thereof
  • Channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and construction method thereof
  • Channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and construction method thereof

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experiment example

[0019] The first step, construction and screening of recombinant lentivirus:

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Abstract

The invention discloses a channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line and a construction method of the channelrhodopsin-2-green fluorescence protein (GFP) gene engineered nerve stem cell line. The cell line is the recombined nerve stem cell line C 17.2 of a channelrhodopsin-2 (ChR2) which is capable of stably expressing and a green fluorescence protein (GFP). The recombined ChR2-GFP gene engineered nerve stem cell line C 17.2 is obtained by constructing recombined slow virus, transducing C 17.2 nerve stem cell line and sifting. The channelrhodopsin-2 (ChR2)-green fluorescence protein (GFP) gene engineered nerve stem cell line can be used for in vivo and in vitro studies on the functional integration of a nerve neuron of a transplanted stem cell differentiation source and a nervous system of a host and provides a favorable platform for the study on the functional integration of a nerve neuron of a transplanted stem cell differentiation and a nervous system of a host. Especially with the utilization of a filter paper digestion method, the stem cell recombining method improves the rate of GFP positive cells, is beneficial to streaming Fluorescence Activated Cell Sorting (FACS) separation, and greatly improves the efficiency of an experiment.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a ChR2-GFP genetically engineered neural stem cell line and a construction method thereof. Background technique [0002] Nervous system degenerative diseases are a class of diseases, including Parkinson's, Alzheimer's and other diseases. At present, there is no effective and specific therapy, and stem cell transplantation alternative therapy has a good prospect. The neurons derived from stem cells transplanted into the body must be able to establish functional synaptic connections with the remaining neurons of the host nervous system in order to restore the transmission of synaptic information. The current various studies only provide indirect evidence in terms of morphology, but have not yet clarified the issue functionally, especially at the level of synapses that transmit information. The development of optogenetics technology can solve this problem. The use of genetic technology ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/867
Inventor 王少军
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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