Difference between revisions of "Part:BBa K3502011"

 
Line 3: Line 3:
 
<partinfo>BBa_K3502011 short</partinfo>
 
<partinfo>BBa_K3502011 short</partinfo>
  
−
This year, we tested the editing efficiency of ADAR1 by adding a pTRE system and a stem ring. The essence grains were added with fluorescent protein after the PTRE promoter to report the editing efficiency. We found that the addition of the stem ring inhibited the background leak of the PTRE promoter and had no significant effect on its peak value, which we thought might be an important pTRE improvement.
+
This year, we tested the editing efficiency of ADAR1 by adding a pTRE system and a stem ring. The essence grains were added with fluorescent protein after the pTRE promoter to report the editing efficiency. In this system, it is hoped that ADAR1 will edit the editing sites on the stem ring to suppress downstream toxic proteins expression, which is replaced by GFP
 +
<br style="clear: both" />
 +
[[File:T--SYSU-CHINA--contri-p4.png|500px|thumb|left]]
 +
<br style="clear: both" />
 +
 
 +
<br style="clear: both" />
 +
In order to know how many DOX our system needs to work effectively, and how the stem-loop function to this system. we do an experiment about the relation between the concentration of DOX and GFP expression, which shows the induced expression efficiency of pTRE was demonstrated,
 +
[[File:T--SYSU-CHINA--contri-p1.png|500px|thumb|left]]
 +
<br style="clear: both" />
 +
And we use a STEM-LOOP effectively reduce the local expression of pTRE.
 +
<br style="clear: both" />
 +
[[File:T--SYSU-CHINA--contri-p2.png|500px|thumb|left]]
 +
<br style="clear: both" />
 +
 
 +
We transferred the plasmid contains STEM-Loop into HeLa cells (with high endogenous ADAR1 expression), and noticed that plasmid with stem ring was effectively inhibited when Dox was low, indicating the success of ADAR1 in stem ring editing.Since the inhibitory effect of ADAR1 was relieved at high Dox expression, we established the specification for experiments at low DOX-induced concentration
 +
<br style="clear: both" />
 +
[[File:T--SYSU-CHINA--contri-p1.png|500px|thumb|left]]
 +
<br style="clear: both" />
 +
And we use a STEM-LOOP effectively reduce the local expression of pTRE.
 +
<br style="clear: both" />
 +
[[File:T--SYSU-CHINA--contri-p2.png|500px|thumb|left]]
 +
<br style="clear: both" />
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 03:08, 28 October 2020


a GFP-STEM-LOOP-apoptin vector controlled by TRE-on system

This year, we tested the editing efficiency of ADAR1 by adding a pTRE system and a stem ring. The essence grains were added with fluorescent protein after the pTRE promoter to report the editing efficiency. In this system, it is hoped that ADAR1 will edit the editing sites on the stem ring to suppress downstream toxic proteins expression, which is replaced by GFP

T--SYSU-CHINA--contri-p4.png



In order to know how many DOX our system needs to work effectively, and how the stem-loop function to this system. we do an experiment about the relation between the concentration of DOX and GFP expression, which shows the induced expression efficiency of pTRE was demonstrated,

T--SYSU-CHINA--contri-p1.png


And we use a STEM-LOOP effectively reduce the local expression of pTRE.

T--SYSU-CHINA--contri-p2.png


We transferred the plasmid contains STEM-Loop into HeLa cells (with high endogenous ADAR1 expression), and noticed that plasmid with stem ring was effectively inhibited when Dox was low, indicating the success of ADAR1 in stem ring editing.Since the inhibitory effect of ADAR1 was relieved at high Dox expression, we established the specification for experiments at low DOX-induced concentration

T--SYSU-CHINA--contri-p1.png


And we use a STEM-LOOP effectively reduce the local expression of pTRE.

T--SYSU-CHINA--contri-p2.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1348
    Illegal BamHI site found at 10
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1015