Part:BBa_K3638003
pEGFP-miR-155-sponge-1 sensor
the GFP value of cells transfected with pEGFP-miR-155-sponge-1 plasmid could show the different expression of miRNAs in cells
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1028
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
result of BBa_K3638003
effect of pEGFP-miR-155-sponge-1 in breast cancer cells
MicroRNA-155 (miR-155) is a well-known oncogenic miRNA overexpressed in many human cancers, including breast cancer[5]. So we constructed pEGFP-miR-155-sponge-1 and try to test the possibility of detecting miR-155 expression by using this plasmid. In order to explore whether XIST correlates with pEGFP-miR-155 sensor in breast cancer cells with potential target sites. pEGFP–C1 (as negative controls), pEGFP-miR-155-sponge-1 (0.8 ug plasmids for each well) was transfected into human breast cells (MDA-MB 231 cells and MDA-MB 468 cells) in 24-well plate, respectively. After transfection, cells were examined under fluorescence microscopy (Fig.4 A, B, C, D and Table 1). The fluorescence of GFP was decreased in MDA-MB 231 cells and MDA-MB 468 cells transfected with pEGFP-miR-155-sponge-1 compared with controls (Fig. 5). In addition, we also measured the value of GFP fluorescence by plate reader (SpectraMax i3). The endogenous miR- 155 could inhibit the expression of GFP in cells transfected with pEGFP-miR-155-sponge-1 (Table 1 and Fig5). The results suggested the GFP value of cells transfected with pEGFP-miR-155-sponge-1 could show the different expression of miRNAs in cells.
(A). pEGFP–C1 was transfected in MDA-MB 468 cells. (B). pEGFP-miR-155-sponge-1 was transfected in MDA-MB 468 cells. (C). pEGFP–C1 was transfected in MDA-MB 231 cells. (D).pEGFP-miR-155-sponge-1 was transfected in MDA-MB 231 cells.
Different cells were transfected with pEGFP-C1 or pEGFP-miR-155-sponge-1 for 48 h.
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