A kind of preparation method of dna or rna complex

A complex and cationic polymer technology, applied in the field of biomedical materials, can solve the problems of unfavorable DNA or RNA transfer, poor controllability of complex structure, large size difference, etc., and achieve strong repeatability of operation and reproducible size Strong control and uniform size effect

Inactive Publication Date: 2011-12-07
PEKING UNIV
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AI Technical Summary

Problems solved by technology

[0004] At present, the method for preparing complexes by mixing DNA or RNA with cationic polymers is mainly simple mixing. The mixing methods include vortex mixing, ultrasonic mixing, etc. The complexes prepared by this method have the following disadvantages: (1) The complexes are relatively small in size. Large, usually around 200-500nm, the larger size of the complex is not conducive to the delivery of DNA or RNA; (2) The controllability of the complex structure is poor, and the traditional simple mixing complex process is largely controlled by kinetics, and the formed The structure of the complex is affected by many factors such as concentration, salinity, mixing order, mixing speed, etc. The structure and morphology of the formed complex largely depend on the preparation process, and the size difference between the prepared complex particles is small. Large, not conducive to delivery of DNA or RNA

Method used

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  • A kind of preparation method of dna or rna complex
  • A kind of preparation method of dna or rna complex
  • A kind of preparation method of dna or rna complex

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Experimental program
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Effect test

Embodiment 1

[0054] Dilute the standard phosphate buffer solution 5 times to prepare 0.2×PBS buffer solution; dissolve the DNA in the buffer solution at a concentration of 5×10 -4 g / ml; Polylysine (PLL) was dissolved in buffer at a concentration of 1×10 -3 g / ml; using a capillary electrophoresis instrument to provide an electric field; inject DNA and PLL from both ends of the capillary successively with pressure. In the presence of an electric field, DNA flows to the positive electrode, and PLL flows to the negative electrode. DNA and PLL flow in the presence of an electric field Mix down to obtain DNA or RNA complexes. Fix the injection volume of DNA, adjust the voltage and the injection volume of PLL to obtain DNA or RNA complexes of different sizes.

[0055] An atomic force microscope AFM (NANOSCOPE IIIa, EV scanning head, tapping mode) was used to observe the structure of the complex prepared under the electric field. The size of the complex was uniform, and the size was about 100 nm....

Embodiment 2

[0057] Dilute the standard phosphate buffer solution 5 times to prepare 0.2×PBS buffer solution; dissolve RNA in the buffer solution at a concentration of 5×10 -5 g / ml; Polylysine (PLL) was dissolved in buffer at a concentration of 1×10 -4 g / ml; using a capillary electrophoresis apparatus to provide an electric field; inject RNA and PLL from both ends of the capillary successively with pressure. In the presence of an electric field, RNA flows to the positive pole, and PLL flows to the negative pole. RNA and PLL flow in the presence of an electric field. Mix down to obtain RNA complexes. Fix the injection volume of RNA, adjust the voltage and the injection volume of PLL, and obtain RNA complexes of different sizes.

[0058] An atomic force microscope AFM (NANOSCOPE IIIa, EV scanning head, tapping mode) was used to observe the structure of the complex prepared under the electric field. The size of the complex was uniform, and the size was about 100 nm.

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Abstract

The invention discloses a method for preparing a DNA (Deoxyribonucleic Acid) or RNA (Ribonucleic Acid) complex, belonging to the field of biological medicinal materials. In the method, by applying an external direct current electric field, a cationic polymer solution and a DNA or RNA solution meet each other in the electric field and are compounded to form a complex of DNA or RNA and cationic polymer. The DNA or RNA complex prepared by the method has a controllable structure and a small and uniform size and contributes to transfer of DNA or RNA and gene therapy.

Description

technical field [0001] The invention relates to a preparation method of a DNA or RNA complex, in particular to a preparation method of a DNA or RNA complex with controllable size and good repeatability, and belongs to the field of biomedical materials. Background technique [0002] In recent years, with the in-depth study of DNA, scientists have discovered that many diseases, such as genetic diseases and tumors, are closely related to human genetic abnormalities. Gene therapy is an innovative biomedical technology developed in the 1990s. It introduces human normal genes or genes with therapeutic effects into human target cells in a certain way to correct gene defects or exert therapeutic effects, so as to achieve the purpose of treating diseases. Gene therapy can be used to treat many genetic diseases such as hemophilia and cystic fibrosis, as well as malignant diseases such as cancer and AIDS. In 1990, American scientists successfully carried out human gene therapy for pa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87
Inventor 梁德海杨旭燕牛林夏玉琼刘赟郑萃周继寒孙建波苏翠翠
Owner PEKING UNIV
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