A method for regulating cell pathways by using plant hormone aba and small molecular substance pyr

A technology of small molecular substances and plant hormones, applied in the field of biological method invention, can solve problems such as incompatibility, immune system damage, etc., and achieve the effect of reducing damage

Inactive Publication Date: 2021-08-03
JILIN UNIV
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  • Abstract
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Problems solved by technology

However, both this method and optogenetics have certain limitations
Rapamycin is an immunosuppressor, which has certain damage to the immune system; the optogenetics system is not suitable for the regulation of intracellular light signaling pathways

Method used

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  • A method for regulating cell pathways by using plant hormone aba and small molecular substance pyr
  • A method for regulating cell pathways by using plant hormone aba and small molecular substance pyr
  • A method for regulating cell pathways by using plant hormone aba and small molecular substance pyr

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Embodiment 1

[0040] A method for regulating the Wnt signaling pathway in human HEK293T cells by using the plant hormone ABA and the small molecular substance PYR, comprising the following methods:

[0041] (1) Carrier design and construction:

[0042] First utilize the primer described in claim 1, take Arabidopsis thaliana cDNA and HEK293T cell cDNA as template, utilize pcr technology to amplify each gene fragment, then utilize corresponding primer, use two sections of fragment as template, carry out overlapPCR; ABI1 and PYR1 were fused together with the C-terminus of LRP6, a key protein in the Wnt signaling pathway, and PCR products such as ABI1, PYR1, ABI1-LRP6C, and PYL1-LRP6C were recovered using agarose gel, and the endonuclease XbaI XmaI double enzyme Cut the linearized pCIi 4xmyc, pCI egfp, and pCI mcherry vectors, use the Infusion method to connect, transform DH5a, colony PCR verification, pick clones and send them for sequencing, and ensure the correct sequencing; using the abov...

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Abstract

The invention provides a method for regulating cell pathways by using plant hormone ABA and small molecular substance PYR. The ABA receptor proteins ABI1 and PYR1 in Arabidopsis thaliana were constructed on human cell expression vectors, and then transferred into HEK293T cells respectively. Adding ABA, the two receptor proteins will have an inductive interaction. After adding the inhibitor PYR, the interaction The degree of operation will be reduced. After the key proteins of other human cell signaling pathways are fused with these two receptor proteins, the two receptor proteins interact under the induction of ABA, resulting in ABA-induced aggregation of the fused signaling pathway. The present invention selects the key protein LRP6 on the human Wnt pathway. After the fusion expression of ABA receptor and LRP6, LRP6 will aggregate under the induction of ABA, thereby promoting the production of β-catenin and opening the intracellular signaling pathway. After adding PYR inhibition, the production of β-catenin decreases and the cellular pathway is closed.

Description

technical field [0001] The invention relates to the field of biological method inventions, specifically, a method for regulating cell pathways by using plant hormone ABA and small molecular substance PYR. Background technique [0002] Optogenetics (Optogenetics) mainly uses photoreceptors that can undergo multimerization after receiving light induction or photoreceptors that can undergo photospecific interactions with their interacting proteins, and then couple related signal transduction proteins to Regulates cell signal transduction. So far, many photoreceptors have been improved to fuse key proteins in signaling pathways to regulate and control many pathways in organisms, such as receptor tyrosine kinase Ras-MAPK signaling pathway, PI3K-Akt signaling pathway and Wnt signaling pathway. At the gene level, the researchers used elements such as CRY2-CIB1, FKF1-GI, UVR8-COP1, and LOV proteins to regulate the transcription and expression of specific genes in mammalian cells. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N5/10
CPCC12N15/85C12N2840/002
Inventor 左泽乘张力莫伟亮张峻川李苏迪石翔肖勇张文礼张宝昌
Owner JILIN UNIV
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