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Composition and method for generating linear polymer through light-operated RAFT polymerization in living cells

A linear polymer and composition technology, applied in the field of cell biology, can solve the problems of small polymer molecular weight and uncontrollable reaction, and achieve the effect of low molecular weight distribution and low cytotoxicity

Active Publication Date: 2021-10-12
SHENZHEN INST OF ADVANCED TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, due to the high activity of the free radical reaction itself, most of the produced polymers have a wide molecular weight distribution (the reaction is uncontrollable), and due to the complex chemical environment in the cell, the highly active free radical intermediates are easy to react with amino, sulfhydryl Chain transfer occurs through the action of active groups, so the molecular weight of the obtained polymer is relatively small

Method used

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  • Composition and method for generating linear polymer through light-operated RAFT polymerization in living cells
  • Composition and method for generating linear polymer through light-operated RAFT polymerization in living cells
  • Composition and method for generating linear polymer through light-operated RAFT polymerization in living cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1: the synthetic steps of RAFT chain transfer agent 2-(butylthiocarbonylthiothiolthio)propionic acid

[0078] Such as figure 2 As shown, within 30min, add 50mL THF solution containing 16.0g n-butanol dropwise to 150mL THF suspension containing 9.0g KOH, stir at room temperature for 30min, then add dropwise 50mL solution containing 17.0g CS 2 After stirring at room temperature for 24 hours, the THF solution was concentrated to 50 mL under reduced pressure. Under stirring, 50 mL of THF solution containing 22.4 g of n-propylammonium bromide was added dropwise to the above concentrated solution. After stirring at room temperature for 24 hours, the solvent was removed under reduced pressure to obtain the target crude product. The product was further purified by silica gel column chromatography to obtain the target product as bright yellow crystals.

Embodiment 2

[0079] Example 2: Synthesis steps of biotin-modified acrylate monomer BiotinMA

[0080] Such as Figure 4 As shown, 1.0g biotin, 1.5g poly(ethylene glycol) methacrylate (Mn=500Da), 0.78g N-(3-dimethylaminopropyl)-N'-ethylcarbodiethylene Amine hydrochloride and 0.05g 4-(dimethylamino)pyridine were dissolved in 50mL DMF, stirred at room temperature for 24h, and the solvent was removed under reduced pressure. 3 , 1% hydrochloric acid and saturated brine for 3 times, dried with anhydrous sodium sulfate, and removed dichloromethane under reduced pressure to obtain the target crude product, which was further purified by silica gel column chromatography to obtain the target product as a white viscous liquid.

Embodiment 3

[0081] Example 3: CCK-8 test characterizes cell viability

[0082] In the CCK-8 test, HeLa cells were first treated with 1x10 4 The density was inoculated in a 96-well cell culture plate and incubated at 37°C and 4% carbon dioxide for 18 hours at a constant temperature to ensure that the cells adhered to the wall; 2 ), and continue to incubate at constant temperature for 24 hours; after 24 hours, suck out the original medium and wash the cells with PBS for 3 times, add 100 μL of diluted CCK-8 solution (ratio to medium: 1:10) to each well, incubate at constant temperature for 4 hours and use The microplate reader detects the ultraviolet absorption at 450nm in each well, and compares the absorbance of the treated cells with the untreated cells to obtain the cell activity value.

[0083] In the above method, through the CCK-8 test, it is proved that 10min of light and the composition will hardly cause toxicity to living cells within the working concentration range, as shown in t...

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Abstract

The invention discloses a composition and a method for generating a linear polymer through light-operated RAFT polymerization in living cells. A monomer, a chain transfer agent, a photoinitiator and living cells are incubated together to achieve uptake of the composition by the cells, and then in-situ synthesis of the polymer in the cells is achieved through illumination. The polymer synthesized in the cells is further extracted, represented, tested and compared with a polymer obtained through in-vitro simulation, and it is found that the polymer synthesized in the cells has high molecular weight and narrow molecular weight distribution and belongs to typical polymer products with controllable polymerization. Therefore, the purpose of introducing the unnatural polymer with a controllable structure into the living cell can be achieved, the composition and the structure in the cell can be further artificially regulated, and even the purpose of controlling the cell behavior and the cell function can be possibly achieved.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a composition and a method for generating linear polymers by light-controlled RAFT polymerization in living cells. Background technique [0002] Due to the high complexity of the intracellular chemical environment and the sensitivity of living cells to external stimuli, many organic synthesis reactions are difficult to carry out in cells without damaging cells, so the currently known artificial chemicals that can be carried out in living cells The number of responses is limited. However, the artificial synthesis of unnatural compounds in living cells has great potential in the field of biomedicine, and has always been an important research field in biological research. It is expected to realize the synthesis of artificial organelles by artificially synthesizing unnatural compounds in cells. , regulation of cell structure (such as viscosity, cytoskeleton structure, etc.), an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08F283/06C08F220/54C08F2/38C08F2/48C12N5/09
CPCC08F283/065C08F2/38C08F2/48C12N5/0682C12N5/0693C08F2438/03C12N2529/10C08F220/54Y02A50/30
Inventor 耿晋张一川高权连前进何荣坤
Owner SHENZHEN INST OF ADVANCED TECH
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