Part:BBa_K897318

Paired termini structure

As we know, all antisense RNAs had a short half-life of about 1 minute. Because antisense RNA efficiency is determined by the binding rate to the target, which in turn is determined by both antisense RNA concentration and binding rate constant, a higher intracellular concentration of the inhibitor will result in higher efficacy.

An alternative strategy to stabilize asRNAs is to pair the termini using flanking inverted repeats to create a hairpin structure with the antisense sequence within a large loop. A paired termini (PT) design, where flanking inverted repeats create paired asRNA termini, was proved can produce effective gene silencing. PT asRNAs are abundant and stable and function through an RNase III independent mechanism that requires a large stoichiometric excess of asRNA. PT consists of non-endogenous GC-rich sequences and has a stem-loop structure, which can improve the stability of as RNA raises its abundance.

Meanwhile, the paired termini structure include several multiple clone sites between two stem-loop sequences, as a result, different kinds of antisense fragments can be implanted into this kit.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 59
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 59
  Illegal NotI site found at 84
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 59
  Illegal BamHI site found at 53
  Illegal XhoI site found at 93
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 59
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 59
  
• 1000
  COMPATIBLE WITH RFC[1000]