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Cell surface engineering method for controlling intercellular interaction and application of method

A cell surface and cell technology, applied in the field of controlling the interaction between cells, can solve the problem of no decomposition control, etc., and achieve the effect of migration

Active Publication Date: 2021-02-26
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, tunable disassembly of aggregated cellular assemblies remains a challenge, as most existing studies have not focused on disaggregation control

Method used

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  • Cell surface engineering method for controlling intercellular interaction and application of method
  • Cell surface engineering method for controlling intercellular interaction and application of method
  • Cell surface engineering method for controlling intercellular interaction and application of method

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Experimental program
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Effect test

Embodiment 1

[0029] (1) Before constructing the DNAzyme-based logic gate, we use a Zn 2+ The specific RNAcleavingDNAzyme (strand-1) and its substrate strand (strand-2) act as control switches, manipulating cellular assembly and disassembly. In the following cell assembly experiments, the cells were divided into two groups. Such as figure 1 In a, these two groups of cells were modified with strand-1 and strand-2, respectively. Both chains are modified with a cholesterol at the 5' end, allowing insertion into the cell membrane. The 3' ends of strand 1 and strand 2 were fluorescently imaged with FAM and Cy5, respectively. Cell-to-cell assembly can be achieved by hybridization of -1 and -2 strands. in Zn 2+ In its presence, the substrate-2 strand is cleaved from the ribonucleotide cleavage site, resulting in immediate cell disassembly. Taking HeLa cells as an example, cell coupling and depolymerization under the control of DNAzyme strategy were successfully obtained ( figure 1 b).

[0...

Embodiment 2

[0039] To explore the utility of DNAzyme-controlled intercellular interactions in biological applications, we further used Zn 2+ Specific DNAzymes and their substrates control the assembly and dissociation between two cell spheroids. Using previous DNAzyme / substrate strategies, cell-to-cell attachment and separation can be achieved not only in a single isolated cell, but also in two multicellular spheroids. A Zn observed under the microscope 2+ T-cell spheroids modified with a specific DNAzyme (chain-1) meet and ligate with HeLa tumor cell spheroids modified with a corresponding substrate (chain2). Join Zn 2+ After ionization, the double sphere assembly of T cells / tumor cells can be dissociated. In addition, we have further achieved the migration of T cell spheroids (chain-1 modification) from one tumor cell spheroid (chain-2 modification) to another tumor cell spheroid. When a T-cell spheroid approaches a tumor cell spheroid, the two spheroids form a link through double-s...

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Abstract

The invention provides a cell surface engineering method for controlling intercellular interaction. The method comprises the following steps: respectively modifying the same ends of base sequences ofDNAzyme and a corresponding substrate with long ester chains, respectively modifying to-be-controlled cells with the DNAzyme and the corresponding substrate through the long ester chains, realizing intercellular aggregation by combining the DNAzyme with the corresponding substrate, and adding metal ions corresponding to DNAzyme to cause breakage of the corresponding substrate so as to realize celldissociation. The cell surface engineering method has the advantages that: (1) the DNAzyme and the substrate lipid modified terminal are allowed to be fixed to a cell membrane, and based on the characteristics of the DNAzyme and the substrate, intercellular dynamic controllable regulation among multiple cell spheres is realized; and (2) the T cell spheres can be closer to HeLa cell spheres, and migration of the T cell spheres is realized, so that T cells are promoted to better attack tumor cells, and the application potential of the T cell spheres serving as a new treatment tool in cancer treatment is shown.

Description

technical field [0001] The invention relates to the technical field of cell surface engineering, in particular to a method for controlling interaction between cells and its application. Background technique [0002] Cell membranes are highly complex biological boundaries composed of lipids, proteins, and polysaccharides that protect intracellular material from the environment. Acting as a protective wall, the cell membrane is a multifunctional platform that houses many essential proteins and ligands. Therefore, cell functions and cell behaviors are expected to be regulated through cell surface engineering. At present, the modification of cell membrane has been successfully applied to intercellular coupling, interaction and tissue engineering. Since the binding of ligands and receptors can induce host-guest interactions between cells, bottom-up assembly of cells is usually achieved by modifying the cell membrane with recognizable ligands and receptors. The most widely used...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/09C12N5/071
CPCC12N5/0006C12N5/0636C12N5/0693C12N5/0682Y02A50/30
Inventor 钱若灿周泽蕊陆艺
Owner EAST CHINA UNIV OF SCI & TECH
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