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Application of 2-pentanone and specific receptor thereof in preparation of products for regulating and controlling cell functions

A specific, pentanone technology, applied in receptors/cell surface antigens/cell surface determinants, applications, medical preparations containing active ingredients, etc., can solve the problem of insufficient control accuracy, unsuitable long-term use, Complex operation and other problems, to achieve the effect of rapid onset of effect, good clinical application prospects, and simple experimental operation

Inactive Publication Date: 2022-05-03
湖北优尼泰睿生物科技开发有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] For this reason, the embodiment of the present invention provides a kind of 2-pentanone acceptor and its application, to solve the traditional optogenetics and chemical genetics in the prior art The control technology of cell physiological activities by technical means has problems such as unsuitable for long-term use, complicated operation, inconvenient use, and insufficient control accuracy.

Method used

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  • Application of 2-pentanone and specific receptor thereof in preparation of products for regulating and controlling cell functions
  • Application of 2-pentanone and specific receptor thereof in preparation of products for regulating and controlling cell functions
  • Application of 2-pentanone and specific receptor thereof in preparation of products for regulating and controlling cell functions

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Embodiment 1

[0026] Example 1, Construction of 2-pentanone-specific receptor Or35a and specific receptor 83b expression vectors

[0027] like figure 1 As shown, the specific receptor of 2-pentanone in the embodiment of the present invention can be expressed in the target area of ​​the animal by virus vector or transgenic technology, and 2-pentanone reaches the cells in the receptor expression area by inhalation, and the receptor Binding, opening ligand-gated cation channels, causing intracellular calcium ion concentration to increase, cell membrane depolarization, and electrical or endocrine activities.

[0028] The construction method of the 2-pentanone-specific receptor Or35a and specific receptor 83b expression vectors of this embodiment comprises the following steps:

[0029] 1. Construction of Or35a-P2A-Or83b-P2A co-expression gene

[0030] 1. Taking the extracted Drosophila olfactory bulb cDNA as a template, wherein the nucleotide sequence of the Or35a-specific receptor gene is sho...

Embodiment 2

[0047] Embodiment 2, transgenic model animal construction

[0048] This embodiment provides the construction process of transgenic model animals as follows:

[0049] 1. Construction of homologous recombination vector: use the CRISPR / Cas9 approach to construct a homologous recombination vector, place the exogenous gene Or35a-P2A-Or83b-P2A fragment between the two homologous arms of the Rosa26 gene, and select CAG for extensive activation son. After a series of enzyme digestion and identification, the homology of the vector reached 100% after sequencing, and it can be confirmed that the targeting vector was constructed successfully.

[0050] Construction and identification of sgRNA vectors: (1) Synthesize gRNA primers according to the Rosa26 site sequence; (2) Phosphorylate and anneal single-stranded oligo to form double strands, denature at 98°C, cool to room temperature to form DNA with sticky ends Fragment; (3) Plasmid PX459 was cut with BbsⅠ, recovered by gel cutting after...

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Abstract

The embodiment of the invention discloses application of 2-pentanone and a specific receptor thereof in the following (A1) or (A2): (A1) preparing a product for regulating and controlling cell functions; (A2) regulating and controlling cell functions; (A3) preparing a product for promoting the increase of the content of calcium in cells; and (A4) preparing a product for promoting the increase of the discharge frequency of neurons. According to the invention, a 2-pentanone specific receptor is expressed in a cultured cell or animal body, and through specific binding with 2-pentanone, a ligand gating cation channel is opened, and in the process of causing intracellular calcium ion concentration increase, cell membrane depolarization, electrical activity or endocrine generation and other activities, accurate regulation and control of tissue cell and organ functions are finally realized. After treatment, the cells can be quickly activated to generate effects, and the effects are quickly taken; after the treatment is stopped, the experiment effect can be quickly terminated, and the original state is recovered; the cells cannot be damaged after long-time treatment, and long-time treatment can be realized; the method is convenient to transform to clinical application and is used for disease treatment.

Description

technical field [0001] The embodiment of the present invention relates to the field of biotechnology, and specifically relates to the application of 2-pentanone and its specific receptor in the preparation of products regulating cell functions. Background technique [0002] In the field of basic research in life sciences, neurons, cardiomyocytes, skeletal muscle cells, etc. have the ability to emit and transmit action potentials, and have important physiological functions in the life process. Researchers usually use some means to activate neurons in a specific area of ​​the brain, or stimulate cardiomyocytes, skeletal muscle cells, and intracellular calcium ion-dependent physiological functions (such as cell secretion processes) to stimulate a specific behavior or produce Some important physiological functions should be studied in depth. The application of such technologies can play an important role in the treatment of major diseases such as psychiatric diseases, nervous s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K31/121A61P25/00A61P9/00A61P5/00C07K14/705C12N15/12C12N15/85C12N5/10
CPCA61K38/177A61K31/121A61P25/00A61P9/00A61P5/00C07K14/705C12N15/85C12N2740/16043A01K2217/07A01K2227/105C12N15/907A01K2267/03C12N15/86C07C49/20C12N2502/00
Inventor 李斌
Owner 湖北优尼泰睿生物科技开发有限公司
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