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Gene control device and method based on CRISPR-Cas9

A technology of gene regulation and one nucleotide, applied in the field of genes

Active Publication Date: 2017-01-11
THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, limited by complex and diverse biological signals, CRISPR-Cas9 technology cannot ideally modify the endogenous gene network

Method used

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  • Gene control device and method based on CRISPR-Cas9
  • Gene control device and method based on CRISPR-Cas9

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Embodiment Construction

[0020] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0021] figure 1 A schematic diagram showing the working principle of the method for regulating gene expression in one embodiment, including an overview of the recombinant sgRNA silencing target gene (a) or activating the target gene (b), the sgRNA structurally includes two parts: and the inactivation Cas9 protein (dCas9) binding first part (M) and the second part (N) connected to the 3' end of the first part (M), wherein the 5' end of the first part (M) has an antisense sequence (M1), the second Portion (N) is an aptamer sequence portion comprising a ligand binding portion (N1) and an aptamer stem portion (N2). In the absence of signal A (or B), that is, when there is no ligand, the antisense sequence (M1) of the sgRNA is paired with the aptamer stem (N2), and the sgRNA is in a "closed" state; In the case of A (or B), that is, w...

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Abstract

The invention discloses a gene control device and method based on CRISPR-Cas9. The device comprises a nucleotide sequence, wherein the nucleotide sequence is an sgRNA itself or can be transcribed into an sgRNA, the sgRNA structurally comprises a first part bonded with inactivated Cas9 protein and a second part connected to the 3'end of the first part, the 5'end of the first part is provided with an antisense sequence, and the second part is an aptamer sequence part which comprises a ligand binding part and an aptamer stem part; when no ligand exists, the aptamer sequence is pair-bonded with the aptamer stem part; when a ligand exists, the aptamer sequence is dissociated from the aptamer stem part so as to be pair-bonded with a target DNA. By integrating an RNA riboswitch bonded with a specific biological signal into the sgRNA, a bridge of cell signal intensity and gene expression intensity is established, any target gene can be activated or silenced on the basis that biological signal identification is achieved, and artificial connection is built between signal ligands and cytogene.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to a CRISPR-Cas9-based gene regulation device and gene regulation method. Background technique [0002] Eukaryotic cells have complex biological behaviors, such as proliferation, apoptosis, migration and differentiation, which are closely regulated by cell signaling pathways and genetic networks. The key node in the transformation of cell signaling pathways is to establish a close relationship between specific input signals and output signals. In order to better understand the mechanism of different signaling pathways to achieve complex biological responses, synthetic biologists have begun to design various engineered cells with novel signaling pathways. Recent related research has made a good elaboration and exploration on the construction of new sensors and signal processing systems, but such signal processing systems cannot ideally link the detection of specific signals with the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85
Inventor 黄卫人刘宇辰蔡志明陈志聪
Owner THE SECOND PEOPLES HOSPITAL OF SHENZHEN
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