Chitosan as a Non-viral Gene Delivery System for hcp Gene of Campylobacter jejuni

Sarannya, E (2017) Chitosan as a Non-viral Gene Delivery System for hcp Gene of Campylobacter jejuni. Masters thesis, Indian Institute of Science Education and Research, Kolkata.

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Abstract

delivery holds potential solution for a plethora of disorders and diseases such as SCID, diabetes, cancer haemophilia and many more. Its immense application in the gene therapy and vaccine development necessitate the need for a suitable delivery platform that can overcome the problems associated with conventional vector based delivery strategy. To this end, Chitosan, a cationic smart polymer, out-weights vector based strategy in many ways. Chitosan is known to be biocompatible, biodegradable, pH responsive and cost effective. Moreover, the pH sensitive nature of chitosan makes it a suitable candidate for gene delivery via mucosal route. Although chitosan have been used in the field of drug and gene delivery for decades, the mechanism of its pH responsiveness to facilitate the targeted delivery of cargos is not clearly understood. The efficacy of chitosan based nanoparticles as a gene delivery vehicle has already been reported in the past. The F gene delivery of New castle disease virus, gene therapy based on factor VIII for hemophilia, Gonadotropin releasing hormone for cancer therapy are few of them. With an aim to assess the potential of chitosan based nanoparticles in gene delivery in the present study we chose to use hcp gene of Campylobacter jejuni. C.jejuni is considered to be leading cause of food borne infections. Recent reports showing the presence of Type VI secretion system (T6SS) has revolutionized the mode C.jejuni infection in human and chicken. Hemolysin Coregualted Protein is an integral part of T6SS and may aid in bacterial pathogenicity. However, co-localization and its active target in host cells are yet to be determined. In order to accomplish this, we have constructed mammalian expression vector (pEGFPN1) encoding hcp gene of C.jejuni isolated from human clinical samples. Subsequently chitosan based nano-particle encapsulating pEGFPN1-HCP construct was prepared using complex coacervation method. Finally, pH and temperature dependent release of hcp gene from Chitosan-nanoparticles and the biocompatible nature of CS nanoparticles were ascertained. Further study to assess the in-vivo co-localization of hcp gene delivered via CS nanoparticle is suggested.

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