Integrated synchronous deoxyribonucleic acid (DNA) nano device, application of integrated synchronous DNA nano device to living cell multi-target imaging, and living cell multi-target imaging method of integrated synchronous DNA nano device
A nano-device and living cell technology, applied in the field of live cell imaging, can solve the problems of limited reaction rate, different module entry efficiency, signal difference, etc., and achieve the effect of improving imaging accuracy, simple reaction system, and avoiding movement differences
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Embodiment 1
[0036] Example 1: Imaging of single target miR-21 in living cells by using integrated synchronous DNA nanodevices of different designs.
[0037] Using the HeLa cell line of cervical cancer cells as the basic model, 20 μL of integrated synchronous DNA nanodevice probes were added to HeLa cells pre-cultured in eight-well confocal culture dishes (0.2 mL, 1×10 6 mL -1), incubated at 37°C for 4 hours. After washing with PBS, imaging was performed using a confocal laser fluorescence microscope. In the target miR-21-positive cells, miR-21 replaces the Block chain that seals the R-SD and releases the single-stranded R-SD, whose 5' end 14 bases are connected to the loop corresponding to the R-HD chain The 14 bases in the shape part are complementary to form a double strand, and the damaged base oG site on the R-HD chain can be acted on by the enzyme hOGG1 to form an apurinic / apyrimidinic AP site, which is then digested by APE 1 to release FAM fluorescence.
[0038] In addition, the...
Embodiment 2
[0040] Example 2: Using an integrated synchronous DNA nano-device to simultaneously image multiple targets in different living cells.
[0041] Using the HeLa cell line of cervical cancer cells as the basic model, 20 μL of integrated synchronous DNA nanodevice probes were added to HeLa cells pre-cultured in eight-well confocal culture dishes (0.2 mL, 1×10 6 mL -1 ), incubated at 37°C for 4 hours. After washing with PBS, imaging was performed using a confocal laser fluorescence microscope. In the presence of the target miR-21, the strand displacement reaction and the Block DNA strand complement each other to form a double strand, releasing the R-SD strand; the T-SD-S strand, the primer at the 5' end is extended in the presence of the target telomerase T-SD chain. The 14 bases at the 5' end of the R-SD chain, T-SD chain and C-SD chain are complementary to the 14 bases of the ring part of the corresponding R-HD chain, T-HD chain and C-HD chain to form a double chain; After the...
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