Part:BBa_K779314

mKate-HHm - Repressible hammerhead behind mKate MammoBlock

Along with Part:BBa_K779311, Part:BBa_K779312, and Part:BBa_K779313, this part introduces in vivo hammerhead ribozymes to iGEM and the registry. Hammerhead ribozymes are self-cutting strands of catalytic RNA. After a hammerhead ribozyme is transcribed in a cell, it folds into a secondary structure, which causes self-cleavage. Because of their RNA-cleaving function, hammerheads may be used to regulate protein expression and to make other RNA parts. For more information on hammerheads in general, see http://en.wikipedia.org/wiki/Hammerhead_ribozyme .

These four parts consist of a hammerhead ribozyme attached to an mKate florescent protein for reporting purposes. The two non-repressible hammerheads are designed to show that hammerheads can self-cleave in a mammalian environment. After the hammerhead-mKate sequence is transcribed, the mRNA sequence should cleave, preventing the transcription of mKate. Successful cleavage can be verified by comparing fluorescence levels to a constitutive mKate control. The two repressible hammerheads are designed to test an antisense strategy for interfering with hammerhead function. An interfering RNA should be able to bind to the modified repressible hammerheads, preventing the hammerhead secondary structure from forming and leaving the transcript intact. This can be observed in an increase in fluorescence, when compared to the regular hammerheads.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]