Introduction: Multidrug resistance bacteria is alarming worldwide. A lot of research were done and are ongoing to search for the best, convenient and economically affordable ways to fight them. With the latest genome editing tool; Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology, this research was performed to develop a novel strategy to genetically modify the genome and inhibit the growth of Klebsiella pneumoniae (UPM ESBLKP1), an Extended Spectrum Beta Lactamases (ESBL) organism. Methods: A CRISPR-Cas9 vector was construct- ed together with guide RNAs designed specifically for the targeted uppP gene, a gene responsible for bacterial cell growth and protection. Results: The growth and cell wall integrity of the modified Klebsiella pneumoniae (UPM ESBLKP1) were significantly inhibited and reduced, respectively. Interestingly, wild type Klebsiella pneumoniae showed a normal growth curve while modified strains showed a faster doubling rate when supplemented with Luria-Bertani media. In contrast, slower growth rate of modified strain was observed in the M9 minimal media. This explained the higher doubling rate of mutants on nutrient rich medium earlier is being related to gene recovery. They grew slowly in the minimal media as they were adapting to a new environment while recovering the uppP gene and surviving, proving the success of its gene modification. Conclusion: The developed CRISPR-gRNA system was able to modify the targeted Klebsiella pneumoniae gene hence providing an opportunity to develop a new drug for Klebsiella pneumoniae infection as an alternative to antibiotics.
Cervical cancer is one of the leading causal cancer-related fatalities in the world. Cervical cancer patients can be treated by conventional treatment such as surgery, radiotherapy, chemotherapy, medications and combination treatments. Currently, more targeted treatments are being developed to cure cervical cancer. The treatments include immunotherapy, virotherapy and gene therapy which will be discussed in this paper. In immunotherapy, the synergy of CTLA-4 suppression and PD-1/PDL-1 immune checkpoint inhibition targeting their corresponding pathways enhanced the human immune system resulting a promising treatment effects. Oncolytic viruses such as Newcastle disease virus selectively infect and kill cancerous cells/tissues without harming normal cells/tissues. This character has made them a potential modality in combating cancer which popularly known as oncolytic virotherapy. Gene therapy delivers modified genetic materials to the target cancer cells via viral and non-viral vectors. It is used to target the abnormal gene, to increase cells’ susceptibility towards drugs or conventional therapy, to induce tumour cells apoptosis, to enhance tumour cell immunogenicity recognition and to inhibit the oncogene expression. The objective of this minireview is to add to the general knowledge on aforementioned therapeutic strategies against cervical cancer.