Nature research paper: Cas12a2 elicits abortive infection through RNA-triggered destruction of dsDNA
of around 0.1 were used.BL21 cells were sequentially electroporated with the nuclease-encoding and target/non-target plasmids. TheCas12a-expressing plasmids pCBS273 and pCBS3588 were used, respectively. Target plasmid pCBS273 and non-target plasmid pCBS3578 were used. For the experiments involvingCas13a, nuclease-expression plasmid pCBS361 was used in combination with the target plasmid pCBS2004 and non-target plasmid pCBS612.
The resulting data were analysed in Python. First, clusters of bacteria exhibiting distinct FSC and Pacific Blue signals were identified using density-based spatial clustering of applications with noise . Next, the ratios of the Pacific Blue to the FSC signal for each data point and the percentage of the data points within each cluster were parsed from the clustering data. The resulting values were plotted in the form of balloon plots. A total of 60,000 events per sample were analysed.
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