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Team:SYSU-China/Project/Results
From 2013.igem.org
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In order to test the devices, elements were cloned into our two major plasmids backbone: pcDNA3.0 and p199.The maps or design are shown in each parts of Results. We drove the Suicide Gene with CMV promoter to ensure a robust expression and prominent phenotype in cells. An eGFP was used to indicate the performance of Tet-off and MicroRNA-Target system, which could be quantitated through Image J and Western-Blot. All separately testing experiments were carried out via transient transfection in Bosc and HepG2, considering both the transfection efficiency and representation of liver cancer cell. Also, a survival experiment was done on liver cell to ensure that its miR122 level can successfully knockdown Suicide Gene with miR122 target-site.</p> | In order to test the devices, elements were cloned into our two major plasmids backbone: pcDNA3.0 and p199.The maps or design are shown in each parts of Results. We drove the Suicide Gene with CMV promoter to ensure a robust expression and prominent phenotype in cells. An eGFP was used to indicate the performance of Tet-off and MicroRNA-Target system, which could be quantitated through Image J and Western-Blot. All separately testing experiments were carried out via transient transfection in Bosc and HepG2, considering both the transfection efficiency and representation of liver cancer cell. Also, a survival experiment was done on liver cell to ensure that its miR122 level can successfully knockdown Suicide Gene with miR122 target-site.</p> | ||
| - | <p><strong>If you want to see the results, please click the following wheer gears.</strong></p> | + | <p><strong>If you want to see the results for testing each parts, please click the following wheer gears.</strong></p> |
<br /><img src=" https://static.igem.org/mediawiki/2013/archive/d/db/20130927131402%21Result-overview.png " width="700" /><br /> | <br /><img src=" https://static.igem.org/mediawiki/2013/archive/d/db/20130927131402%21Result-overview.png " width="700" /><br /> | ||
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After confirmation of each device, we proceeded to working on iPSC and assembly of the whole pathway. The whole pathway was integrated into two p199 plasmids, one for regulating and one for response .Lenti-virus packaging these two plasmids was produced and 3 cell lines of iPSC, HepG2, Hela were transfected. This successfully gave us the stable cell line with our iPSC Safeguard design. Then further test and characterization of the pathway’s performance of working as a whole were carried out. All the data can be seen in >>>>>. | After confirmation of each device, we proceeded to working on iPSC and assembly of the whole pathway. The whole pathway was integrated into two p199 plasmids, one for regulating and one for response .Lenti-virus packaging these two plasmids was produced and 3 cell lines of iPSC, HepG2, Hela were transfected. This successfully gave us the stable cell line with our iPSC Safeguard design. Then further test and characterization of the pathway’s performance of working as a whole were carried out. All the data can be seen in >>>>>. | ||
</p> | </p> | ||
| - | <p><strong>If you want to see the results, please click the following cute cells.</strong></p> | + | <p><strong>If you want to see the results testing iPSCs in different periods, please click the following cute cells.</strong></p> |
<br /><img src=" https://static.igem.org/mediawiki/2013/d/db/Result-overview.png " width="700" /><br /> | <br /><img src=" https://static.igem.org/mediawiki/2013/d/db/Result-overview.png " width="700" /><br /> | ||
Revision as of 13:32, 27 September 2013
ipsc
Overview of result
In order to test the devices, elements were cloned into our two major plasmids backbone: pcDNA3.0 and p199.The maps or design are shown in each parts of Results. We drove the Suicide Gene with CMV promoter to ensure a robust expression and prominent phenotype in cells. An eGFP was used to indicate the performance of Tet-off and MicroRNA-Target system, which could be quantitated through Image J and Western-Blot. All separately testing experiments were carried out via transient transfection in Bosc and HepG2, considering both the transfection efficiency and representation of liver cancer cell. Also, a survival experiment was done on liver cell to ensure that its miR122 level can successfully knockdown Suicide Gene with miR122 target-site.
If you want to see the results for testing each parts, please click the following wheer gears.
After confirmation of each device, we proceeded to working on iPSC and assembly of the whole pathway. The whole pathway was integrated into two p199 plasmids, one for regulating and one for response .Lenti-virus packaging these two plasmids was produced and 3 cell lines of iPSC, HepG2, Hela were transfected. This successfully gave us the stable cell line with our iPSC Safeguard design. Then further test and characterization of the pathway’s performance of working as a whole were carried out. All the data can be seen in >>>>>.
If you want to see the results testing iPSCs in different periods, please click the following cute cells.
Sun Yat-Sen University, Guangzhou, China
Address: 135# Xingang Rd.(W.), Haizhu Guangzhou, P.R.China
