Team:Shenzhen/Project/YAO.Suicider
From 2012.igem.org
(Difference between revisions)
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<h5>Original Ideas</h5> | <h5>Original Ideas</h5> | ||
<ul><p>Originally, we try to construct such an artificial organelle YAO that YAO receives signal either from biosensor (internal) or from cytoplasm (external) or consequently trigger the host cell apoptosis. However, for YAO are not completed, we use mitochondrion as experiment target. And since we can’t rely on the availability of YAO.Sensor, the idea that Initial signal is given to YAO’s promoter is replaced by that nucleus produce T7 RNAP and transport it into mitochondrion to activate the T7 promoter inside it.</p></ul> | <ul><p>Originally, we try to construct such an artificial organelle YAO that YAO receives signal either from biosensor (internal) or from cytoplasm (external) or consequently trigger the host cell apoptosis. However, for YAO are not completed, we use mitochondrion as experiment target. And since we can’t rely on the availability of YAO.Sensor, the idea that Initial signal is given to YAO’s promoter is replaced by that nucleus produce T7 RNAP and transport it into mitochondrion to activate the T7 promoter inside it.</p></ul> | ||
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p1.jpg]] | [[File:project_yao_suicider_p1.jpg]] | ||
- | <p>Figure 1. Original ideas.</p></div> | + | <p>Figure 1. Original ideas.</p></div></ul> |
</div> | </div> | ||
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<ul><li>f. Apoptosis of Host</li></ul> | <ul><li>f. Apoptosis of Host</li></ul> | ||
<ul><p>Apoptosis factor cytochrome C and largely fleeing calcium can trigger the caspase-like pathway. Aif1 such as endo G can.get into nucleus and fragmentate DNA. Finally, host cell is suppressed or even die.</p></ul> | <ul><p>Apoptosis factor cytochrome C and largely fleeing calcium can trigger the caspase-like pathway. Aif1 such as endo G can.get into nucleus and fragmentate DNA. Finally, host cell is suppressed or even die.</p></ul> | ||
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p2.jpg]] | [[File:project_yao_suicider_p2.jpg]] | ||
- | <p>Figure 2. Final stories.</p></div> | + | <p>Figure 2. Final stories.</p></div></ul> |
</div> | </div> | ||
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<h5>Verification Experiments Design</h5> | <h5>Verification Experiments Design</h5> | ||
<ul><li>For T7 Imitation System</li></ul> | <ul><li>For T7 Imitation System</li></ul> | ||
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p3.jpg]] | [[File:project_yao_suicider_p3.jpg]] | ||
- | <p>Figure 3. Verify the function of T7 RNAP/Promoter system in cytoplasm. If it works, fluorescence of GFP can be detected in cytoplasm.</p></div> | + | <p>Figure 3. Verify the function of T7 RNAP/Promoter system in cytoplasm. If it works, fluorescence of GFP can be detected in cytoplasm.</p></div></ul> |
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p4.jpg]] | [[File:project_yao_suicider_p4.jpg]] | ||
- | <p>Figure 4. Verify the function of T7 RNAP/Promoter system in mitochondrion or unfinished YAO, basing on the availability of signal peptide from cytoplasm to mitochondrial matrix. If it works, fluorescence of mtGFP can be detected in mitochondrion.</p></div> | + | <p>Figure 4. Verify the function of T7 RNAP/Promoter system in mitochondrion or unfinished YAO, basing on the availability of signal peptide from cytoplasm to mitochondrial matrix. If it works, fluorescence of mtGFP can be detected in mitochondrion.</p></div></ul> |
<ul><li>For Retrograde Signaling Part</li></ul> | <ul><li>For Retrograde Signaling Part</li></ul> | ||
<ul><p>Because DLD3 is one of the target of RTG pathway of yeast, lowering the basal expression is required.</p></ul> | <ul><p>Because DLD3 is one of the target of RTG pathway of yeast, lowering the basal expression is required.</p></ul> | ||
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p5.jpg]] | [[File:project_yao_suicider_p5.jpg]] | ||
- | <p>Figure 5. Verify the basal expression of gene downstream DLD3 promoter. Roughly judge the brightness of GFP, and consequently select a suitable degradation tag for gene downstream DLD3 promotor.</p></div> | + | <p>Figure 5. Verify the basal expression of gene downstream DLD3 promoter. Roughly judge the brightness of GFP, and consequently select a suitable degradation tag for gene downstream DLD3 promotor.</p></div></ul> |
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p6.jpg]] | [[File:project_yao_suicider_p6.jpg]] | ||
- | <p>Figure 6. Verify the function of selected degradation tag by using UV as a imitation of DNase produced in mitochondrion that can lead to mitochondrion dysfunction which will drive the gene expression downstream DLD3 promoter through Retrograde signaling from mitochondrion to nucleus.</p></div> | + | <p>Figure 6. Verify the function of selected degradation tag by using UV as a imitation of DNase produced in mitochondrion that can lead to mitochondrion dysfunction which will drive the gene expression downstream DLD3 promoter through Retrograde signaling from mitochondrion to nucleus.</p></div></ul> |
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p7.jpg]] | [[File:project_yao_suicider_p7.jpg]] | ||
<p>Figure 7. Verify that holing can make pores on the inner membrane of mitochondrion. This is the alterant of DNase causing mt dysfunction. And it can also produce a membrane without mtDNA.</p></div> | <p>Figure 7. Verify that holing can make pores on the inner membrane of mitochondrion. This is the alterant of DNase causing mt dysfunction. And it can also produce a membrane without mtDNA.</p></div> | ||
<ul><li>For Host Cell Suicide Part</li></ul> | <ul><li>For Host Cell Suicide Part</li></ul> | ||
- | <div class="figurep"> | + | <ul><div class="figurep"> |
[[File:project_yao_suicider_p8.jpg]] | [[File:project_yao_suicider_p8.jpg]] | ||
- | <p>Figure 8. Verify the function of holing in yeast to prove that holing can act as bax/bak protein in human cell as a apoptosis activator. If it works, we can see that between the engineered and versus one, the growthrate curve (OD600) of engineered one tend to be lower than that of versus one.</p></div> | + | <p>Figure 8. Verify the function of holing in yeast to prove that holing can act as bax/bak protein in human cell as a apoptosis activator. If it works, we can see that between the engineered and versus one, the growthrate curve (OD600) of engineered one tend to be lower than that of versus one.</p></div></ul> |
</div> | </div> | ||
</div> | </div> | ||
{{:Team:Shenzhen/Temp/gallery.htm}} | {{:Team:Shenzhen/Temp/gallery.htm}} |
Revision as of 13:47, 25 September 2012