Method and its Composition for encapsulation, stabilization, and delivery of siRNA in Anionic polymeric nanoplex: An In vitro- In vivo Assessment

Abstract

Small interfering RNA (siRNA) are synthetic RNA duplex designed to specifically knockdown the abnormal gene to treat a disease at cellular and molecular levels. In spite of their high potency, specificity, and therapeutic potential, the full-fledged utility of siRNA is predominantly limited to in vitro set-up. Till date, Onpattro is the only USFDA approved siRNA therapeutics available in the clinic. The lack of a reliable in vivo siRNA delivery carrier remains a foremost obstacle towards the clinical translation of siRNA therapeutics. To address the obstacles associated with siRNA delivery, we tested a dendrimer-templated polymeric approach involving a USFDA approved carrier (albumin) for in vitro as well as in vivo delivery of siRNA. The developed approach is simple in application, enhances the serum stability, avoids in vivo RNase-degradation and mediates cytosolic delivery of siRNA following the endosomal escape process. The successful in vitro and in vivo delivery of siRNA, as well as targeted gene knockdown potential, was demonstrated by HDAC4 inhibition in vitro diabetic nephropathy (DN) podocyte model as well as in vivo DN C57BL/6 mice model. The developed approach has been tested using HDAC4 siRNA as a model therapeutics, while the application can also be extended to other gene therapeutics including micro RNA (miRNA), plasmids oligonucleotides, etc.

Effect of Cationic Lipid Nanoparticle Loaded siRNA with Stearylamine against Chikungunya Virus

“…The ODA-based lipoplex (F4) was prepared at different (N/P) atomic ratios that were tested: 1, 2, 4, 6, 8 and 10, keeping the siRNA quantity constant at 1000 pmol. The lipid nanoparticle size and charge were observed using a Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) [45].…”

“…The obtained pellets were re-suspended in PBS and acidic solutions (3 mL). Further, the pellets were stirred at 100 rpm for 7 days at 37 • C. At different time points, the samples were aliquoted and were centrifuged at 13,000× g for 28 min at 25 • C [45]. The collected supernatant with an equal volume of fresh PBS or acidic solution was added to the sample.…”

Development and Characterization of Ultrasound Activated Lipopolyplexes for Enhanced Transfection by Low Frequency Ultrasound in In Vitro Tumor Model

“…The anionic surface charge of UCVs maintains its vascular retention for a longer period. [ 3,31 ] The prepared ultrasound contrast vesicles exhibited 70% equivalent gray‐scale ultrasound contrast as compared to SonoVue (Figure 2a). It was observed that DPPC/PEG40S (95:5) formed a foamy suspension due to immiscibility of amphiphilic PEG40S in lipid dispersion after the agitation process.…”

Immunonano-Lipocarrier-Mediated Liver Sinusoidal Endothelial Cell-Specific RUNX1 Inhibition Impedes Immune Cell Infiltration and Hepatic Inflammation in Murine Model of NASH

“…This given dose to the animal was determined through the confirmation of encapsulation of siRNA as per the protocol reported by Cun and Raval with slight modifications [36,37]. For the %EE determination, accurately weighed (5.0 mg) lyophilized particles matrix were solubilized in 100 µL of chloroform (for SC shell) with 400 µL of aqueous TE buffer (for SPC core).…”