Preparation method and application of blue 4',6-diamidino-2-phenylindole dye tracing gene nano-carrier
A nano-carrier, phenylindole technology, applied in the fields of biotechnology and medicine, can solve the problems of inability to determine the target cell exogenous gene, the specific location of the difficult-to-trace gene carrier, etc., and achieve low cytotoxicity and high gene transfection efficiency Effect
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Embodiment 1
[0027] Mesoporous silica gene nanocarrier loaded with DAPI dye and transfected GFP plasmid into Hela cells:
[0028] 1) DAPI dye-embedded mesoporous silica gene nanocarrier: dissolve the gene nanocarrier described in step 1 into 2 ml of deionized water and transfer to a 10 ml glass bottle, then add 10 mg of DAPI The dyestuff was stirred and reacted at 100 rpm at 50°C for 6 hours. After the reaction, centrifuge at 11,000 rpm for 20 minutes, and wash the precipitate once with deionized water to obtain a mesoporous silica gene nanocarrier that can track exogenous genes in real time.
[0029] 2) Adsorption of plasmid DNA by the mesoporous silica gene nanocarrier: Dissolve the mesoporous silica gene nanocarrier capable of real-time tracking of exogenous genes obtained in the above step 1) into 100 microliters of deionized water. Take a sterile 1.5 ml centrifuge tube, add 50 μl of serum-free medium OPTI-MEM and 0.3 μg of green fluorescent protein (GFP) plasmid DNA and mix well, the...
Embodiment 2
[0032] Mesoporous silica gene nanocarrier loaded with DAPI dye and transfected GFP plasmid into Hela cells:
[0033] 1) DAPI dye-embedded mesoporous silica gene nanocarrier: dissolve the gene nanocarrier described in step 1 into 3 milliliters of deionized water and transfer to a 10 milliliter glass bottle, then add 30 milligrams of DAPI Dye, stirred and reacted at 300 rpm at 35°C for 9 hours. Immediately after the reaction, centrifuge at 10,000 rpm for 15 minutes, and wash the precipitate twice with deionized water to obtain a mesoporous silica gene nanocarrier that can track exogenous genes in real time.
[0034]2) Adsorption of plasmid DNA by mesoporous silica gene nanocarriers: the mesoporous silica gene nanocarriers obtained in the above step 1) that can track exogenous genes in real time were dissolved in 300 microliters of deionized water. Take a sterile 1.5 ml centrifuge tube, add 100 μl of serum-free medium OPTI-MEM and 1 μg of green fluorescent protein (GFP) plasmid ...
Embodiment 3
[0037] Mesoporous silica gene nanocarrier loaded with DAPI dye and transfected GFP plasmid into Hela cells:
[0038] 1) DAPI dye-embedded mesoporous silica gene nanocarrier: Dissolve the gene nanocarrier described in step 1 into 5 ml of deionized water and transfer to a 10 ml glass bottle, then add 50 mg of DAPI Dye, stirred and reacted at 500°C for 12 hours at 500 rpm. Immediately after the reaction, centrifuge at 11,000 rpm for 20 minutes, and wash the precipitate three times with deionized water to obtain a mesoporous silica gene nanocarrier that can track exogenous genes in real time.
[0039] 2) Adsorption of plasmid DNA by the mesoporous silica gene nanocarrier: Dissolve the mesoporous silica gene nanocarrier capable of real-time tracking of exogenous genes obtained in the above step 1) into 500 microliters of deionized water. Take a sterile 1.5 ml centrifuge tube, add 200 μl of serum-free medium OPTI-MEM and 3 μg of green fluorescent protein (GFP) plasmid DNA and mix w...
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