Difference between revisions of "Part:BBa K4586025"
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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. | 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. | 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. | ||
Revision as of 10:37, 21 September 2023
Cargo (guide RNA - switch- Cas12k- MCP/ADAR)
Description
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.
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