Part:BBa_K2514000
pSB1C3-miR-21-sponge-6s
Our project constructed a mir-21 sponge containing marker gene of GFP for the detection and inhibition mir-21 in the cell lines, and offers a non-invasive and highly sensitive approach for early diagnosis and treatment of colorectal cancer in the future. miRNA sponge, which contains multiple target sites complementary to a miRNA of interest, is used to be a inhibitor of miRNA. Here, we proposed miRNA sponge could be a monitor to reflect the change of miRNA expression in cells. After transfection of the miR-21 sponge, sponge RNAs can attract miR-21, inhibiting the expression of eGFP reporter. The expression of miR-21 should be monitored by measuring the expression of eGFP reporter.Pervious studies indicated sponges containing bulged sites that are mispaired opposite miRNA are more effective. Typical sponge constructs contain 2 to 10 binding sites separated by a few nucleotides each. Here, we designed two different mir-21 sponges with different binding sits to test the efficiency of sponges. One miR-21 sponge contains two miR-21 binding sites with 3-nt spacers for bulged sites. Another one mir-21 sponge contains six miR-21 binding sites with 3-nt spacers for bulged sites. We wanted to compare the accuracy and the effectiveness of different miR-21 sponges contained different binding sites for measuring the expression of miR-21 in colorectal cancer cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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