CRISPR-Cas9 Gene Editing in Bacteria: lacZ Gene Modification

Overview

CRISPR-Cas9 leads to the alteration of the genetic code, allowing for in-depth disease research and improvements in daily function with humans, animals, and even plants. But how does this biotechnology do this? What allows for precise cutting to implement the new genetic code? How can we ensure this process works effectively? This study hypothesizes that the CRISPR-Cas9 system enables gene code alteration, as evidenced by phenotypic and genotypic results. The Escherichia coli (E. coli wild-type strain) was extracted from the control group (IX/ARA) plate, the plate with the donor guide and arabinose, as well as the plate with the donor guide lacking arabinose. These plates were used to conduct polymerase chain reaction testing, amplifying our DNA and preparing it for gel electrophoresis to illustrate how the various band lengths and sizes determine whether the process had successfully (genotypically) worked. Following gel electrophoresis, the data analysis showed genotypic variations as expected. Phenotypic results showed different band lengths between the blue (unedited) and white (edited) colonies. Blue colonies represented bands at 1100 bp and 350 bp, consistent with the wild-type lacZ gene, while white colonies lacked the 1100 bp band but possessed bands at 680 bp and 350 bp. This indicated the successful insertion of the donor DNA. These results support the hypothesis that CRISPR-Cas9 accurately worked to alter the Escherichia coli’s genetic code, producing white colonies rather than blue.