Nano-heparin sodium-pei-ca2+ gene transfer material and preparation method
A technology of gene introduction and -pei-ca2, which is applied in the field of functionalized nano-heparin sodium-PEI-Ca2+ gene introduction materials and preparation, can solve the problems of limited penetration of the gene gun and unstable results, and achieves good biocompatibility Good performance, good material dispersion, good repeatability
Inactive Publication Date: 2018-08-03
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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[0018] 1. Preparation of the gene transfer material of the present invention:
[0019] 1. Material: Main materials: calcium chloride, polyetherimide (PEI), sodium heparin, analytical grade, Aldrich product; use without further purification steps, Aldrich product. All glassware for preparation was sonicated in ethanol for 5 minutes, then washed with double distilled water, and washed with lotion H 2 0 / HNO 3 (65%) / H 2 O 2 (35%) (1:1:1, v / v / v) cleaning, followed by double-distilled water, acetone, and then dried in the air.
[0020] 2. Material preparation: Dissolve calcium chloride in deionized water to prepare 0.1M freshly prepared calcium chloride solution; dilute the heparin sodium solution with deionized water to prepare 2g / L freshly prepared heparin sodium solution; prepare 4g / L freshly prepared PEI solution . First, add 4mL of 2g / L freshly prepared heparin sodium solution, +1mL of 4g / L freshly prepared PEI solution, and +2mL of 0.1M freshly prepared calcium chloride solution t...
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Abstract
The invention discloses a nano-heparin sodium-PEI-Ca2+ gene introduction material and a preparation method thereof, which comprises heparin sodium, polyetherimide (PEI) and calcium chloride to form heparin sodium-PEI-PEI with an average diameter of about 100nm Ca2+ nano-spherical material, the content of heparin sodium is 0.54g / g, the content of polyetherimide (PEI) is 0.16g / g, and the content of calcium ion is 0.30g / g. The invention also discloses a preparation method of the nano-heparin sodium-PEI-Ca2+ gene transfer material. The invention has high transfection efficiency, which can reach about 40% in human breast cancer cells; low toxicity, because heparin sodium and calcium chloride have good biocompatibility, and the cell survival rate is high; the material dispersibility is good , meeting the requirements for transfection; very low cost, simple and easy-to-operate material preparation reaction, and good reproducibility.
Description
[Technical Field] [0001] The invention relates to a functionalized nano heparin sodium-PEI-Ca with high transfection efficiency and low cytotoxicity 2+ Gene transfer materials and preparation methods. [Background technique] [0002] With the completion of the human gene mapping, our understanding of the pathogenesis of human diseases has made a breakthrough at the genetic level. Current research shows that there are many diseases that are closely related to genes. If gene fragments or gene mutations that are specifically related to the disease can be screened out, specific treatments can be targeted at the gene level, such as by introducing related missing genes or silencing genes to enhance related missing functions or silencing disease-causing genes , So as to achieve the purpose of thorough treatment. The safe and effective introduction of target genes into organisms is currently the key and difficult point in this research field. [0003] Gene transfer systems or methods can...
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IPC IPC(8): C12N15/85C08L5/10C08L79/08C08K3/16
Inventor 王深明张德元刘勇夏浩明周鸿雁李梓伦常光其徐安武
Owner 王深明