Gene transfer material and preparation method
A technology of gene introduction and gene transfection, applied in the direction of introducing foreign genetic material using vectors, recombinant DNA technology, etc., can solve the problems of limited penetrating power of gene guns and unstable results, and achieve good biocompatibility and low cost , good repeatability
Inactive Publication Date: 2012-07-18
王深明 +2
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Problems solved by technology
Since there are many serious deficiencies in the viral gene transfer system, such as the possibility of activation of proto-oncogenes by viral transfection, the non-viral transfection method is currently a research hotspot. In the above-mentioned non-viral transfection, microinjection once Only one cell can be processed; the penetration of the gene gun is very limited; the results of the calcium phosphate co-precipitation method are unstable; only the cationic liposome method shows good transfection efficiency, but the high toxicity limits its application
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[0015] One, the preparation of the gene transfer material of the present invention (ACC, functionalized nano-amorphous calcium carbonate after calcium phytate modification):
[0016] 1. Raw material preparation
[0017] Analytical pure calcium chloride, ammonium carbonate, phytic acid (molecular weight, MW660), the aforementioned raw materials are all Aldrich company products. All the glassware used for raw material preparation were sonicated in ethanol for 5 minutes, then washed with double distilled water, and rinsed with washing solution H 2 O / HNO 3 (65%) / H 2 o 2 (35%) (1:1:1, v / v / v), then washed with double distilled water and acetone in turn, and finally dried in air.
[0018] 2. Preparation method of gene transfer material
[0019] Dissolve calcium chloride in deionized water to prepare a fresh 0.01M calcium chloride solution. First, add 80mg of phytic acid to 80mL of 0.01M calcium chloride solution, stir and mix well, and divide it into four 25mL beakers , 20mL in...
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Abstract
The invention discloses a gene transfer material, which is a compound material consisting of calcium carbonate and calcium phytate and having the average grain diameter of about 200 nm, wherein the content of the calcium carbonate is 80%, and the content of the calcium phytate is 20%. The invention further discloses a preparation method of a gene transfer material. The gene transfer material has the advantages of higher transfection efficiency reaching about 40% in the human vascular smooth muscle cells, low toxicity and high cell survival because the calcium carbonate has good biocompatibility, good dispersity and suitability of the requirements of the transfection and low cost because the phytic acid is food additive and has low cost. In addition, the chemicals, i.e. calcium chloride and ammonia in the experiment are common cheap reagents; the preparation method of the material is simple and easy to operate and has good repeatability.
Description
technical field [0001] The present invention relates to gene introduction technology. Background technique [0002] Gene transfer systems or methods can be divided into two categories: viral gene transfer systems, using retroviruses, adenoviruses, and adeno-associated viruses as carriers; non-viral gene transfer systems, such as microinjection, gene guns, calcium phosphate co-precipitation, Cationic liposome method, and emerging nano-gene delivery materials. Since there are many serious deficiencies in the viral gene transfer system, such as the possibility of activation of proto-oncogenes by viral transfection, the non-viral transfection method is currently a research hotspot. In the above-mentioned non-viral transfection, microinjection once Only one cell can be processed; the penetration of the gene gun is very limited; the results of the calcium phosphate co-precipitation method are unstable; only the cationic liposome method shows good transfection efficiency, but the ...
Claims
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IPC IPC(8): C12N15/63C12N15/85
Inventor 王深明周鸿雁徐安武叶财盛张德元刘勇胡作军王冕常光其
Owner 王深明