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Transmembrane peptide modified pseudovirus gene vector system

A membrane-penetrating polypeptide and carrier system technology, which is applied in the fields of gene carrier systems and imitation virus gene carrier systems to achieve the effects of improving gene transfection efficiency, simple preparation and efficiency improvement

Inactive Publication Date: 2006-07-12
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are a series of problems such as safety in direct use of viral vectors.

Method used

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  • Transmembrane peptide modified pseudovirus gene vector system
  • Transmembrane peptide modified pseudovirus gene vector system
  • Transmembrane peptide modified pseudovirus gene vector system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Dissolve chitosan in an appropriate amount of 1% acetic acid solution to prepare a 0.2% solution, and dissolve pEGFP-N2 green fluorescent protein particle DNA in an appropriate amount of 50mM sodium sulfate solution to prepare a 100μg / ml solution. Equal volume vortex 30s mixes and makes chitosan-DNA solution, then the homologous protein (abbreviation: Antp) of drosophila transcription factor Antennapedia is dissolved in appropriate amount of 10% acetic acid solution, and after adjusting pH to 7.0 with sodium hydroxide, is prepared into Add 0.1% polypeptide solution and plasmid DNA solution in equal volume to the chitosan-DNA solution, and vortex at 55°C for 30s to mix.

Embodiment 2

[0032] Dissolve chitosan in an appropriate amount of 1% acetic acid solution to prepare a 0.2% solution, and dissolve pEGFP-N2 green fluorescent protein particle DNA in an appropriate amount of 50mM sodium sulfate solution to prepare a 100μg / ml solution. Equal volume vortex for 30s to make chitosan-DNA solution, then dissolve Antp in appropriate amount of 10% acetic acid solution, adjust pH to 7.0 with sodium hydroxide, prepare 0.05% peptide solution, add equal volume to plasmid DNA solution Put it into the chitosan-DNA solution, and vortex at 55°C for 30s to mix well.

Embodiment 3

[0034] Dissolve chitosan in an appropriate amount of 1% acetic acid solution to prepare a 0.2% solution, and dissolve pGL2-ControlVector luciferase plasmid DNA in an appropriate amount of 50mM sodium sulfate solution to prepare a 100μg / ml solution. Equal volume vortex 30s and mix to make chitosan-DNA solution, then dissolve Antp in appropriate amount of 10% acetic acid solution, adjust pH to 7.0 with sodium hydroxide, prepare 0.005% peptide solution, and add in equal volume with plasmid DNA solution Put it into the chitosan-DNA solution, and vortex at 55°C for 30s to mix well.

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Abstract

The invention relates to a film-through polypeptide modified virus imitation gene carrier system in the field of biology technology. It adopts nanometer particle which is formed by high molecular material with film-through polypeptide modified kation to prepare for the virus imitation gene carrier system. It uses the mode of admixing the virus imitation surface and the polypeptide modified non-virus carrier and simulating the virus to enters into the cell to improve the gene transmitting effect.

Description

technical field [0001] The invention belongs to the field of biological technology and relates to a gene carrier system, in particular to a virus-like gene carrier system modified by a membrane-penetrating polypeptide. Background technique [0002] DNA medicine is one of the most eye-catching macromolecular medicines in recent years. The mechanism of action of DNA drugs is different from that of ordinary small molecule drugs. They must successfully cross the cell membrane and nuclear membrane barriers to reach the nucleus before they can begin to encode physiologically active proteins. The cell membrane tightly controls the transport of extracellular substances into the cell, and the barrier function of the cell membrane is the main obstacle that limits the efficiency of DNA expression in cells. At present, in the preparation of DNA drugs, viral and non-viral vectors are mostly used to assist DNA to enter cells. [0003] Although the transfection efficiency of viral vector...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/63C12N15/86C12N15/79C12N15/87C12N15/33
Inventor 蒋晨黄容琴裴元英
Owner FUDAN UNIV
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