Part:BBa_K5242021
gRNA
Our whole system is designed of RNA-sensing in living yeast cells, the gRNA is a simulation of our target transcript, which can form dsRNA with our sensor containing a A-C mismatch. At the appearance of gRNA, the ADAR will recognize the dsRNA and edit A to I at the mismatch point, activating the expression of our payloads (we use eBFP here).
To make sure our sensor RNA expresses with gRNA at the same time, we loaded them in a same plasmid, which is called pSensor1. Also, we set another 2 groups as negative control and 1 group as positive control. In pSensor2, gRNA is mutated, it can bind with sensor RNA perfectly, without a A-C mismatch. We set up this control group to verify the effect of A-C mismatch on ADAR editing. In sensor 3, gRNA is instead by another gRNA, which will not form dsRNA with our sensor RNA. We set up this control group to verify the effect of dsRNA on ADAR editing. In pSensor4, sensor RNA is mutated, the terminal codon UAG was already edited to UGG. We set this control group to check if the 2A peptide and downstream eBFP could work normally.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 48
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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