European Journal of Biomedical and Pharmaceutical Sciences

dnaK GENE EDITING IN THE Escherichia coli GENOME VIA THE CAS9/CRISPR SYSTEM

Mohammad Asaduzzaman*, Farha Matin Juliana, Md. Hayder Ali, Most. Jinnat Parvin, Saad Ibne Hossain, Fazla Rabbi Sharif, Naznin Kazi, Nazia Rifat Zaman and Tasnuva Ferdous

ABSTRACT

The Cas9/CRISPR genetic engineering technology is the centerpiece of a robust adaptive bacterial immune system which contains clustered regularly interspaced palindromic repeats. CRISPR-associated protein-9 (Cas9) is part of this immune system that seeks out, cuts, and degrades viral DNA. By taking advantage of the Cas9/CRISPR system, a tool has been developed to delete or insert DNA into target genes in bacterial cells or mammalian cells to obtain cells with desired traits. In this study, we used the Cas9/CRISPR system for engineering the genome of a non-pathogenic, laboratory strain of bacteria, Escherichia coli. We introduced a deletion mutation in the dnaK gene of the bacteria which is responsible for DNA replication, thus allowing the bacteria to grow on a dnaK template containing a plate. We investigated the dnaK gene deletion efficiency of the Cas9/CRISPR genome editing system.

Keywords: dnaK, Cas9/CRISPR, E. coli.