Preparation method and application of liposome-chitosan compound gene carrier
A technology of gene carrier and chitosan, which is applied in the field of preparation of liposome-chitosan composite gene carrier, can solve problems such as unsatisfactory, increase absorption and bioavailability, solve excessive cytotoxicity, reduce cell toxic effect
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Embodiment 1
[0036] The preparation process adopts a 55 μl complex system and a high-throughput transfection procedure in a 96-well plate. The mass ratio of chitosan and DNA is 1 / 1, the preparation method of 0.1-1 μ g DNA calculation per hole of 96-well plate: 0.1-1 μ g DNA is diluted to 25 μ l with DMEM, 0.3-8 μ g DOTAP (according to DOTAP / DNA=3- The mass ratio of 8 / 1) was diluted to 25 μl with DMEM, and the two were gently mixed (after the liposome dilution was added to the plasmid dilution, it was incubated at room temperature for 5-10min, and 0.1-1μg (according to chitosan / DNA =1 / 1) chitosan, diluted to 5 μ l with DMEM, mixed gently, and incubated at room temperature for 20-30min. That is, the liposome-chitosan composite gene carrier was obtained.
Embodiment 2
[0038] The mass ratio of chitosan and DNA is 2 / 1, the preparation method of 0.1-1 μ g DNA calculation per well of 96-orifice plate: 0.1-1 μ g DNA is diluted to 25 μ l with DMEM, 0.3-8 μ g DOTAP (according to DOTAP / DNA=3- 8 / 1 mass ratio) was diluted to 25 μl with DMEM, after the two were gently mixed (liposome dilution was added to plasmid dilution), room temperature was incubated for 5-10min, and 0.2-2 μg (according to chitosan / DNA=2 / 1) chitosan, diluted to 5 μl with DMEM, mixed gently, and incubated at room temperature for 20-30min. That is, the liposome-chitosan composite gene carrier is prepared.
Embodiment 3
[0040] The mass ratio of chitosan and DNA is 3 / 1, the preparation method of 0.1-1 μ g DNA calculation per well of 96-orifice plate: 0.1-1 μ g DNA is diluted to 25 μ l with DMEM, 0.3-8 μ g DOTAP (according to DOTAP / DNA=3- 8 / 1 mass ratio) was diluted to 25 μ l with DMEM, after the two were gently mixed (liposome dilution was added to the plasmid dilution), room temperature was incubated for 5-10 min, and 0.3-3 μg (according to chitosan / DNA=3 / 1) chitosan, diluted to 5 μl with DMEM, mixed gently, and incubated at room temperature for 20-30min. That is, the liposome-chitosan composite gene carrier is prepared.
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