Difference between revisions of "Part:BBa K1720003"
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We designed 3 silencing device and test their function at the same time.Here is the another two device :BBa_K1720004,BBaK172005 | We designed 3 silencing device and test their function at the same time.Here is the another two device :BBa_K1720004,BBaK172005 | ||
| − | This device with a GFP reporter was then transfected into HEK293 cells by lentiviral vector.Once we silence the PDE5A gene the level of cGMP will be up regulated as a result. The positive control was HEK293 cells that treat with Sodium Nitroprusside ,a NO donator that activate sGC and up regulate the level of cGMP. A negative control was made by transfecting an empty vector that does not contain | + | This device with a GFP reporter was then transfected into HEK293 cells by lentiviral vector.Once we silence the PDE5A gene the level of cGMP will be up regulated as a result. The positive control was HEK293 cells that treat with Sodium Nitroprusside ,a NO donator that activate sGC and up regulate the level of cGMP. A negative control was made by transfecting an empty vector that does not contain scilencing device. We used Elisa to detect cGMP level. The results are as follow: |
[[File:SCUT China shRNA1.png|200px|thumb|left|Green fluorescence signal was observed under fluorescence microscope]][[File:SCUT_China_Elisa_of_scilencing__device.png|200px|thumb|left|cGMP concentration after treat with differenct scilencing device]][[File:SCUT_China_Elisa_of_SNP.png|200px|thumb|left|cGMP concentration after treat with 10umol/L SNP for different time gradient]] | [[File:SCUT China shRNA1.png|200px|thumb|left|Green fluorescence signal was observed under fluorescence microscope]][[File:SCUT_China_Elisa_of_scilencing__device.png|200px|thumb|left|cGMP concentration after treat with differenct scilencing device]][[File:SCUT_China_Elisa_of_SNP.png|200px|thumb|left|cGMP concentration after treat with 10umol/L SNP for different time gradient]] | ||
Revision as of 06:13, 29 August 2015
Human phosphodiesterase 5A gene silencing device NO.1
This device is uesd for silencing the human phosphodiesterase 5A (PDE5A) gene.A U6 promoter driving a designed, synthetic shRNA-like miRNA followed by the terminator.
PDE5A is a cGMP-binding, cGMP-specific phosphodiesterase, a member of the cyclic nucleotide phosphodiesterase family. This phosphodiesterase specifically hydrolyzes cGMP to 5'-GMP. It is involved in the regulation of intracellular concentrations of cyclic nucleotides and is important for smooth muscle relaxation in the cardiovascular system.
We designed 3 silencing device and test their function at the same time.Here is the another two device :BBa_K1720004,BBaK172005
This device with a GFP reporter was then transfected into HEK293 cells by lentiviral vector.Once we silence the PDE5A gene the level of cGMP will be up regulated as a result. The positive control was HEK293 cells that treat with Sodium Nitroprusside ,a NO donator that activate sGC and up regulate the level of cGMP. A negative control was made by transfecting an empty vector that does not contain scilencing device. We used Elisa to detect cGMP level. The results are as follow:
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 273
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 247
- 1000COMPATIBLE WITH RFC[1000]