Part:BBa_K3328005

trigger of OFF-switch2

trigger of 3WJ repressor binding to 3WJ switch(K3328001) to inhibit the expression of downstream gene

Design

3WJ repressor switch RNA employs an unstable hairpin secondary structure. This unstable hairpin was previously demonstrated to be translationally active. When a complementary trigger RNA is expressed, the trigger will bind to the switch RNA, making the originally unstable 3WJ structure stable, and represses translation.

Result

In the validation of 3WJ, as with toehold, we also set blank control (IPTG=0 M, aTc=0 mg/ml). Compared with the group without trigger expression (IPTG=0.1 M, aTc=0 mg/ml), the group with trigger expression (IPTG=0.1 M, aTc=0.25 mg/ml) showed inhibitory effect.

In the validation of 3WJ, as with toehold, we also set blank control (IPTG=0 M, aTc=0 mg/mL). Compared with the group without trigger expression (IPTG=0.1 M, aTc=0 mg/mL), the group with trigger expression (IPTG=0.1 M, aTc=0.25 mg/mL) showed inhibitory effect. Error bar: SD (n=9).

3WJ repressor showed remarkable ON/OFF ratio and had good orthogonality, we choose it as OFF switch. And we used it in the construction of logic gates and the design of some more complex circuits.

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 30
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]