Encapsulation and delivery of plasmid DNA by virus-like nanoparticles engineered from Macrobrachium rosenbergii nodavirus

Nodavirus-based biological container for targeted delivery system

“…Mutations in Asp130 and Asp135 produce intact and broken particles, while mutation of Asp133 disrupts particle formation (Wu et al 2008). We have recently tested the eff ects of reducing and chelating agents on MrNv-VLP stability (Jariyapong et al 2014) and found that this shrimp nodavirus requires both Ca 2 ϩ ions and disulfi de linkages for its assembly process as well as stability of VLP capsid integrity. Interestingly, the assembly of MrNv-VLP selectively depends on calcium ions -only EGTA can disrupt VLP particles in the presence of DTT and not EDTA, which instead causes swelling of particles.…”

Enhancement of shrimp immunity against white spot syndrome virus by Macrobrachium rosenbergii nodavirus-like particle encapsulated VP28 double-stranded RNA

“…We believe that MrNv-VLPs carry VP28 dsRNA into cells by two distinct pathways. Based on an icosahedral structure of MrNV with diameter of 30 ± 3 nm (Goh et al, 2011;Jariyapong et al, 2014), encapsulated VP28 dsRNA-VLP could be readily taken into cells by the macropinocytosis pathway. On the other hand, encapsulated VP28 dsRNA-VLP has a receptor binding region at the protruding domain that facilitates host cell recognition, fusion, and entry to the cell followed by release of VP28 dsRNA into shrimp tissue to act against WSSV Lin et al, 2001).…”

“…Protein expression and purification were carried out according to methods described in our previous work (Jariyapong et al, 2014). Briefly, the pET16b-MrNv constructed with six consecutive histidines was transformed into E. coli BL21(DE3) (New England Biolabs, MA, USA) that was kept at − 80°C and incubated at 37°C overnight, followed by further incubation at 25°C until absorbance reached 0.6-0.8 at 600 nm (A 600 ).…”

“…Disassembly of MrNv-VLP was performed using the protocol described in our previous work (Jariyapong et al, 2014). Briefly, purified MrNv-VLPs were incubated with a disassembling buffer containing 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM ethyleneglycoltetraacetic acid (EGTA), and 20 mM dithiothreitol (DTT) for 1 h at room temperature.…”

Oral delivery of nanoparticle-based vaccines

“…The remaining impurities on VLPs derived from infectious viruses could pose a threat to the safety of vaccines. Moreover, consistent batch-to-batch production of similar sized particles is another hurdle limiting further development of VLPs-based vaccines [96].…”