Part:BBa_K4586025
Cargo (guide RNA - switch- Cas12k- MCP/ADAR)
Description and Usage
The first component of Cargo is the CRISPR system, where we uses guide RNA to direct our Cas12k toward the BAFF-R gene in auto-reactive B-cells, and the second component is the DART V ADAR switch, where we uses ADAR enzyme to convert Adenosine into Inosine, causing a change in the switch condition from off to on status, but ADAR enzyme works only in the presence of complementary mRNA in the target cell to the sensor within the switch that contains the stop codon (UAG) of auto-reactive B-cells. This stop codon contains a mismatched Adenosine (A) group with the complementary mRNA; this group is hybridized by ADAR enzyme activity and converts to an Inosine group (I); thus, the stop codon sequence is disturbed in this condition and our cargo will be translated as shown in figure 1.
Figure 1: This figure illustrates the design of our biological circuit expressing our therapeutic agent under the control of the VP64 transcription module that regulates the activity of the ZF21.16minCMV promoter.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 5988
Illegal AgeI site found at 6100 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1456
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