Liquids in Dishes in a Lab
Small Interfering RNA

RNAi Pathway GraphicThe discovery of RNA interference (RNAi) and the roles that short interfering RNA (siRNA) and micro-RNA (miRNA) play in directing the reversible silencing of genes has enabled the creation of a whole new class of RNA therapeutics to target disease-causing genes that may be otherwise undruggable.

 

RNAi therapeutics currently being developed are poised to revolutionize:

 

• Cancer treatments
• Rare genetic disease treatments
• Immunotherapy

 

 

Small RNA requires a delivery vehicle to protect against nucleases and to facilitate cellular uptake and release into the cytoplasm. Nanoparticles effectively solve several challenges facing small RNA delivery for basic research and drug development:

 

• Protection from nucleases
• Delivery and release into the cytoplasm
• Laborious and time-consuming viral packaging
• Safety concerns with viral vectors
• Toxicity with other delivery methods
• A single approach for RNA delivery for all stages of research and drug development

 

Nanoparticle formulations can be prepared on demand in seconds with NanoAssemblr® technology.


Overcome Challenges in Small RNA Delivery

Several nanoparticle formulations are being explored to package and deliver siRNA including:


Liposome
Liposomes
Polymeric Nanoparticle and Micelles
Polymer NPs and Micelles
Nucleic Acid Lipid Nanoparticle LNP
LNPs

Among these, nucleic acid-LNPs are the most clinically advanced. The first RNAi drug slated for approval (Patisiran) uses LNPs to deliver siRNA. There are however challenges to producing siRNA-loaded nanoparticles that NanoAssemblr® technology addresses:


Challenges with Conventional Methods Benefits of NanoAssemblr® Technology
Significant batch-to-batch variabilityorangeRightArrowCreate highly reproducible siRNA lipid nanoparticles
Substantial material loss from low encapsulation efficiencyorangeRightArrowGenerate siRNA lipid nanoparticles with high encapsulation efficacy and transfection potency
A labor-intensive production process that is difficult to scale up or scale downorangeRightArrowRapidly produce formulations for screening and optimization with a straightforward path to scale-up for clinical applications

Key Benefits


Highly Reproducible siRNA LNP Formulations


siRNA Reproducibility BenchtopParticle size and PDI determined by dynamic light scattering of siRNA LNPs produced by 3 independent users on the NanoAssemblr® Benchtop demonstrating process consistency.


High Encapsulation Efficiency and Potency


siRNA Efficiency & PotencySerum levels of Factor VII protein in mice following i.v. injection of siRNA-LNPs shows a dose-dependent response. At RNA doses of 1 mg/kg over 90% knockdown was achieved for LNPs made on the NanoAssemblr® Benchtop, Blaze, and GMP System.


Straightforward to Scale Production


siRNA Consistent ScalabilityConsistent results for size & PDI of siRNA LNPs achieved across the NanoAssemblr® Platform. Size & PDI were determined by DLS from triplicate formulations. The composition was determined by HPLC.

How It Works
siRNA Production and Delivery
1) An organic solvent containing dissolved lipids and an aqueous solution containing nucleic acids are injected into the two inlet channels of the NanoAssemblr® cartridge.
2) Under laminar flow, the two solutions do not immediately mix, but microscopic features engineered into the channel cause the two fluids to intermingle in a controlled and reproducible way.
3) Within a millisecond, the two fluids are completely mixed, causing a change in solvent polarity that triggers the self-assembly of lipid nanoparticles loaded with nucleic acids.


4) Changing the speed and ratio of fluid injection controls the size of the lipid nanoparticles.
5) Lipid nanoparticles mimic low-density lipoproteins, which allows them to be taken up by an endogenous cellular transport pathway to deliver nucleic acids to cells.
6) Using pH-sensitive lipids allow lipid nanoparticles to release encapsulated nucleic acids into the cytoplasm when vesicle pH decreases.

Get Started

To learn how Precision NanoSystems accelerates nanomedicine development from an idea to clinical applications, contact our Technical Sales Team.

Get in Touch

siRNA Resources

Publication - Summary

May 18, 2019

Annals of Hematology

Lipid nanoparticle-mediated siRNA delivery for safe targeting of human CML in vivo

N. Jyotsana, A. Sharma, A. Chaturvedi, et al.

Read More

Publication - Summary

April 04, 2019

The Journal of Neuroscience

PTCD1 is required for mitochondrial oxidative-phosphorylation: possible genetic association with Alzheimer's disease

D. Fleck, L. Phu, E. Verschueren, T. Hinkle, M. Reichelt, T. Bhangale, B. Haley, Y. Wang, R. Graham, D.S. Kirkpatrick, M...

Read More

Application Note

January 02, 2018

Seamless scale up of siRNA LNP formulations using the NanoAssemblr¬ģ platform

Read More PDF

Publication - Abstract

December 09, 2018

Pharmaceutics

The Microfluidic Technique and the Manufacturing of Polysaccharide Nanoparticles

E. Chiesa, R. Dorati, S. Pisani, B. Conti, G. Bergamini, T. Modena and I. Genta

Read More

Publication - Abstract

December 06, 2018

Advanced Drug Delivery Reviews

Engineering lymphocytes with RNAi

S. Ramishetti, D. Peer

Read More

Publication - Summary

November 21, 2018

Blood

Effective Lipidoid Nanoparticle Delivery In Vivo of siRNA Targeting Kappa Light Chain Production in a Murine Xenograft Model

Xun Ma, Ping Zhou, Adin Kugelmass, Denis Toskic, Melissa Warner, Lisa X. Lee, Teresa Fogaren, Ming Wang, Yamin Li, Liu Y...

Read More
Resource Center

Stay Informed

Sign up today to automatically receive new Precision NanoSystems application notes, conference posters, relevant science publications, and webinar invites.

MENU
X