CRISPR-Cas9 MEDIATED GENOME EDITING IN ESCHERICHIA COLIAbstract
Clustered regularly interspaced palindromic repeats are the centerpiece of a robust adaptive bacterial immune system that forms the basis for the CRISPR-Cas9 genetic engineering technology. CRISPR-associated protein-9 (Cas9) is part of this immune system that seeks out, cuts, and degrades viral DNA. By taking advantage of the CRISPR-Cas9 system, a tool has been developed to delete or insert DNA into target genes in bacterial cells or mammalian cells for obtaining cells with desired traits. In this study, we used the CRISPR-Cas9 system for engineering the genome of a non-pathogenic, laboratory strain of bacteria Escherichia coli. We introduced a mutation in the gene of the bacteria which converted its phenotype from sensitivity to the antibiotic streptomycin to resistance to streptomycin; thus allowing the bacteria to grown on streptomycin containing plates. We also investigated different transformation conditions to determine which condition gave the maximum number of transformants, which reflects the efficiency of the CRISPR-Cas9 genome editing system.
M. Z. Al-Wawi, R. M. Hassan, M. E. Mohamed, M. F. Khan, M. Magaogao and A. Hossain *
Department of Medical Microbiology and Immunology, Ras Al Khaimah Medical and Health Sciences University, Ras Al Khaimah, United Arab Emirates.
27 October 2018
03 February 2019
13 February 2019
01 July 2019