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A kind of functional modification method of cell membrane

A modification method and cell membrane technology, applied in the field of cell engineering, can solve problems such as poor stability, shedding, and reduced ability of probe target recognition.

Active Publication Date: 2021-03-19
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current probes modified by this method have poor stability on the cell membrane and are easily detached from the cell membrane surface in complex biological systems (such as medium containing serum)
In addition, the probes modified on the membrane surface are prone to non-specific interactions with other components, which reduces the target recognition ability of the probes

Method used

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  • A kind of functional modification method of cell membrane
  • A kind of functional modification method of cell membrane
  • A kind of functional modification method of cell membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1: The stability of the amphiphilic tetrahedral probe of the present invention on the cell membrane

[0078] Reduced shedding rate of amphiphilic tetrahedral probes

[0079] After cell counting, 200,000 CEM cells were taken, washed once with PBS, and equally divided into 2 groups, and then mixed with 250 nM S-cho1-FAM (sequence: CCCAGGTTCTCTT- / i6FAMdT / -TTTTTTTTTTTTTT-cholesterol, i6FAMdT represents the place The base T is modified by a fluorescent group) and 250nM T-cho3-FAM prepared by the method of the present invention were co-incubated at 4°C for 10 minutes, washed once with PBS, and dispersed into 1640 medium containing 10% FBS. The two groups of cells were cultured at 37°C in a 5% carbon dioxide environment for 0 hour, 0.5 hour, 1 hour, and 1.5 hour, washed once with PBS, and confocal imaging was used to observe the probe shedding. Such as Figure 6 As shown, the probe S-cho1-FAM was almost completely shed within half an hour, while the probe T-cho3-F...

Embodiment 2

[0080] Example 2: The reduced endocytic efficiency of the amphiphilic tetrahedral probe of the present invention

[0081] After cell counting, 200,000 CEM cells were taken, washed once with PBS, and evenly divided into 2 groups, and then co-incubated with 250nM S-cho1-FAM and 250nM T-cho3-FAM at 4°C for 10 minutes, PBS Wash once and disperse in PBS containing 5 mM magnesium chloride. Both groups of cells were incubated at 37°C for 15 minutes, and the endocytosis of the probe was observed by confocal imaging. Such as Figure 7 As shown, the probe S-cho1-FAM was obviously endocytized after being cultured for 15 minutes, but the probe T-cho3-FAM had no obvious endocytosis. It is revealed that the amphiphilic tetrahedral probe greatly improves the stability on the cell membrane compared with the traditional amphiphilic probe.

Embodiment 3

[0082] Example 3: The amphiphilic tetrahedron probe of the present invention enhances the target recognition ability on the cell membrane

[0083] After cell counting, 200,000 Ramos cells were collected and stained with red live cell tracer (CellTracker TM) stained at 37°C for 15 minutes, washed once with PBS, and divided into 2 groups. Then co-incubated with 250nM T-cho3-sgc8 probe and 250nM S-cho1-sgc8 probe at 4°C for 10 minutes, washed once with PBS, and dispersed into DPBS containing 2% bovine serum albumin (BSA) . Take another 2 million CEM cells, divide them into two groups, add them to the above two groups of Ramos cells, and shake (240rpm) at room temperature for 10 minutes, 20 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 240 minutes, 360 minutes, 540 minutes. The samples at different time points were imaged with a confocal microscope, and the aggregation efficiency of the cell map was counted. Such as Figure 8 As shown, the T-cho3-sgc8 probe-modifi...

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Abstract

The invention discloses a functional modification method for cytomembrane. The method is characterized in that hydrophobic molecules are inserted into a cytomembrane phospholipid bilayer though the hydrophobic effect of the hydrophobic molecules modified on a DNA tetrahedron so as to realize the anchoring of the tetrahedron on the surface of the cytomembrane; a DNA sequence is extended on the vertex, not modified with the hydrophobic molecules, of the DNA tetrahedron, or the extended DNA sequence is combined with other functional units to complete the functional modification of the cytomembrane. The method can not only modify probe molecules on the cytomembrane rapidly, efficiently and losslessly, but also remarkably improve the stability of probes on the cytomembrane and molecular recognition performance of the probes. In addition, based on the programmability of a DNA nanostructure, high-efficient self-assembly of multiple function modules on the cytomembrane can be realized, and a new technology is provided for the research of fields of cell and molecular biology, tissue engineering, cell-based diagnosis and treatment and the like.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a method for functional modification of cell membranes. Background technique [0002] Cell membrane is an important part of eukaryotic cells. It is not only closely related to cell division, proliferation, migration and necrosis, but also plays an important role in the exchange of substances and energy between cells and the external environment. At the same time, cell membranes also regulate cell-cell interactions, cell-cell communication, and intracellular signaling pathways. Therefore, the functional modification of the cell membrane is beneficial to promote basic cell biology research, regulate cell-cell interaction, manipulate cell signal transduction, etc. At present, a large number of functional materials have been modified on the cell membrane, such as DNA, polypeptides, polymer materials and so on. Among these materials, DNA is widely used due to its advantages...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/00
CPCC12N5/0006
Inventor 谭蔚泓邱丽萍李进
Owner HUNAN UNIV
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