Results
Above are results from using a plate reader to measure the OD levels of a 96 well plate. Both experiments (1) and (2) followed the same protocol and antibiotic combinations. Experiment (1) shows the expected results from the new plasmids built in Aim 2. Experiment (2) shows the expected results from Aim 1.
Experiment 2: The samples which were of most relevancy and discussion in the whole picture of our project were the 1229C, 1230C, and 1229F samples. These contained the 1229 or 1230 plasmids. The 1229 plasmid is "special" because when the switch comes in contact with a specific gRNA trigger, the plasmid will "turn off" gRNA activity. The characteristic that we are targeting with 1230 was the “always on".
The 872 plasmid was also transformed into the same electrocompetent cell, with 1229 or 1230. The 872 plasmid includes the ability to cut using Cas12a.
Each of the samples further included a random and correct trigger RNA oligo which in theory should have shut off or activated cell growth – spike in OD measurements. The expected results were as follows.
There was expected growth in the electrocompetent cells that previously had 1229 and were co-transformed with 872 and the control (random) RNA. Similarly, there was not that much growth in the 1230 cells co-transformed with 872 and the control RNA. Importantly, there wasn't growth in the electrocompetent cells that previously had 1229 and were co-transformed with 872 and the experimental RNA. This slight growth is expected behavior from LB after a long duration of time. These results are exciting as they demonstrate the possibility of detecting exogenous RNA with our engineered E. coli.
The rest of our results are shown in the pages for Aim 1, Aim 2 and Aim 3.