Part:BBa_I1012:Design

CI(1) micRNA asRNA

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]

Design Notes

References (unparsed) here:

Mizuno, T., et al. Proc. Natl. Acad. Sci. USA (1984) 81, 1966-1970.

Coleman, J., et al. Cell (1984) 37, 429-36.

References (unparsed) here:

Mizuno, T., et al. Proc. Natl. Acad. Sci. USA (1984) 81, 1966-1970.

Coleman, J., et al. Cell (1984) 37, 429-36.

Complementary to beginning of BBa_I1010 transcript covering RBS, start codon, and 73 bp into coding sequence. No designed secondary structure. Complementary region is flanked by two stem loop structures, as specified by the micRNA mechanism.

Anti-sense

The success of this system clearly rests on the ability to effectively and specifically target mRNA transcripts for degradation using anti-sense RNA. While many papers, articles, and books have been written on the subject, there are no consensus anti-sense building strategies presented. We thus chose to implement three different types of antisense inhibition: KISS, micRNA, and IS10. In the description that follows, the following nomenclature will be used:

target- the mRNA transcript that we wish to inhibit.

anti-sense- the anti-sense molecule which will bind and inhibit target.

micRNA <img src="http://biobricks.ai.mit.edu/IAP_Projects/YoungPower/as_micRNA.gif">

This anti-sense mechanism relies on two stem loops flanking an anti-sense sequence that is specific for the target. The function of the stem loops is to maintain the anti-sense region in a quasi-linear state. BBa_I1012 is built in this manner, with a linear region that will bind over the RBS, start codon, and 76 bp of BBa_I1010.

Incompatible with systems containing BBa_I1011, BBa_I1013.

Compatible with BBa_I1020, BBa_I1021, BBa_I1022, BBa_I1023.

Source

Custom design.