Ontology highlight
ABSTRACT: Background
The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model.Objectives
This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat.Methods
Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR.Results
The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected.Conclusions
PLGA has good potential as a delivery system for pEGFP-C1-tat.
SUBMITTER: Unsunnidhal L
PROVIDER: S-EPMC8636661 | biostudies-literature | 2021 Nov
REPOSITORIES: biostudies-literature
Unsunnidhal Lalu L Wasito Raden R Nugraha Setyawan Erif Maha EM Warsani Ziana Z Kusumawati Asmarani A
Journal of veterinary science 20210830 6
<h4>Background</h4>The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model.<h4>Objectives</h4>This study aims to evaluate the <i>in vitro</i> potential of PLGA as a delivery system for pEGFP-C1-tat.<h4>Methods</h4>Consensus and codon optimization ...[more]