RNA

Part:BBa_I1011:Design

Designed by: June Rhee, Connie Tao, Ty Thomson, Louis Waldman   Group: Antiquity   (2003-01-31)


CI(1) "KISS" asRNA without promoter


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:

Coleman, J., et al. Nature. (1985) 315, 601-3. 

Pestka, S., et al. Proc. Natl. Acad. Sci. USA (1984) 81, 7525-28.

References (unparsed) here:

Coleman, J., et al. Nature. (1985) 315, 601-3. 

Pestka, S., et al. Proc. Natl. Acad. Sci. USA (1984) 81, 7525-28.
Complementary to beginning of BBa_I1010 transcript covering RBS, start codon, and 73 bp into coding sequence. No designed secondary structure.
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.

KISS (Keep it SImple, Silly) <img src="http://biobricks.ai.mit.edu/IAP_Projects/YoungPower/as_KISS.gif">

The simplest of the three methods, this type relies on a single-stranded linear 103 bp anti-sense that is specific to the target of interest. In addition, the first 76 base pairs of the cI region of BBa_I1010 have been codon-modified to give a different sequence that codes for the same cI protein (See BBa_I1030 and I1040).

BBa_I1011 contains the reverse complement of the RBS, start codon, and 76 bp region for BBa_I1010. Thus, if both BBa_I1010 and BBa_I1011 are transcribed, the transcripts will bind to each other and BBa_I1010 will not be translated.

Note that BBa_I1010 already contains a regulatory region, RBS, and coding region (a terminator must be added), while BBa_I1011 does not - thus, when using this component, the appropriate regulatory region, RBS, and terminator must be added to this part.

Incompatible with systems containing BBa_I1012, BBa_I1013.

Compatible with BBa_I1020, BBa_I1021, BBa_I1022, BBa_I1023.



Source

Custom design.

References