02 - Code Breakers: Using CRISPR to Rewrite Genetics
Experiment Results and Analysis
EXPERIMENT 1 Bacteria: E. coli Purple Introduced Plasmid: pCas9-PurpleGuide gRNA/CRISPR Target: purple chromoprotein gene Final Colony Color: White Did CRISPR occur? Yes. The colony color change from purple to white suggests that E. coli Purple went from producing a purple pig- ment to no longer producing a purple pigment. This suggests that the CRISPR-Cas9 system was able to disrupt the purple chromoprotein gene. Total number of colonies: 95
Number of dark purple colonies: 0 Number of light purple colonies: 0 Number of white colonies: 95
EXPERIMENT 2 Bacteria: E. coli Purple Introduced Plasmid: pCas9-ControlGuide gRNA/CRISPR Target: Random sequence not present in E. coli Purple Final Colony Color: Purple Did CRISPR occur? No. The colony color did not change. This suggests that this CRISPR-Cas9 system was not able to disrupt the purple chromoprotein gene responsible for the bacteria’s purple
appearance. This result also confirms that the color change observed in experiment 1 was not solely due to the transformation process (such as an unexpected by-product of intro- ducing the chloramphenicol resistance gene or another gene on the plasmid). Addition- ally, it demonstrates that Cas9 was unable to cut the purple chromoprotein gene without a specifically designed guide RNA. Total number of colonies: 91
Number of dark purple colonies: 91 Number of light purple colonies: 0 Number of white colonies: 0
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