RNA Delivery Technologies: Industrial Applications And Emerging Innovations

By Chloe Tan, Lauren Levy, Yuting Huang, and Ana Jaklenec,* David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology

Recent successes in mRNA vaccines and RNA-based therapies have placed RNA delivery at center stage in biopharma. Encapsulating fragile RNA in protective carriers is essential as naked RNA is rapidly degraded by nucleases in blood and tissues.^1,2 The first FDA-approved RNA drug (patisiran, 2018) used a lipid nanoparticle (LNP) to deliver siRNA,² and the COVID-19 vaccines (approved 2020–21) deployed LNPs in billions of doses worldwide.^1,2 These milestones have spurred massive market growth: the global RNA therapeutics market is projected to grow from about $8.6 billion in 2025 to $26.1 billion by 2034.³ North America represents the leading region for RNA therapeutics (~40% market share),⁴ and LNPs dominate delivery technologies (~41% of market share).⁴ Strategic partnerships (e.g., BioNTech’s 2025 acquisition of CureVac³) and collaborations (e.g., Alnylam’s hypertension program,⁵ BioNTech/CureVac deal⁶) highlight the robust investment into RNA platforms.

The RNA delivery landscape now includes multiple technology classes. LNPs remain the most mature non-viral system, but emerging vectors (exosomes, polymeric nanoparticles, peptide carriers, etc.) are gaining attention. Industry and academic labs are actively tuning carrier chemistry and assembly to address key hurdles: improving stability and endosomal escape, achieving targeted biodistribution, and expanding manufacturing capacity. Below we review the leading approaches and challenges in RNA delivery and outline best practices for formulation and production in RNA therapeutics.

Lipid Nanoparticles

LNPs typically comprise an ionizable lipid, a helper phospholipid, cholesterol, and a PEG-lipid.¹ Upon mixing an RNA-buffer with an ethanolic lipid solution, these components self-assemble into particles that encapsulate the RNA payload.¹

LNPs have become the standard delivery vehicle for mRNA and siRNA drugs due to their high encapsulation efficiency, low immunogenicity, and clinical track record.¹ All FDA-approved LNP therapies (Onpattro siRNA, several mRNA vaccines) use four-component lipids: an ionizable (cationic) lipid to bind RNA, a neutral phospholipid “helper,” cholesterol for membrane structure, and a PEGylated lipid to improve colloidal stability.^1,2 For example, PEG2000-DMG is a common LNP excipient that has been used in formulations for miRNA, siRNA, gene editing, and mRNA vaccines (including COVID-19).⁷ Formulations may also include targeting ligands: LNPs decorated with trivalent galactose–NAcetylgalactosamine (GalNAc) are readily taken up by hepatocytes via the ASGPR receptor,² enabling potent liver-specific delivery (as used in approved siRNA drugs).

Despite their successes, LNPs have limitations. A major challenge is endosomal escape; after endocytosis, only ~2% of internalized LNPs release their RNA into the cytosol.² To address this, developers are designing novel ionizable lipids and helper lipids that disrupt endosomes under acidic conditions.² For example, ionizable lipids with polyamine headgroups (pKa ~6–7) and unsaturated or branched tails can enhance membrane fusion and escape.² Other strategies add peptides or small molecules (e.g., polyhistidine sequences or endosomolytic compounds) to the LNP formulation to promote endosomal release.² Improving escape efficiency can raise potency and reduce required doses — an important consideration, as high doses of LNPs can induce local inflammation.

Another concern is immunogenicity and stability of LNPs. The PEG-lipids that stabilize LNPs can trigger anti-PEG antibodies after repeated dosing, which accelerates LNP clearance and lowers efficacy.² Researchers mitigate this by using lower PEG content, alternative stealth lipids (e.g., zwitterionic PEG mimetics), or by carefully spacing repeat doses. LNPs themselves activate innate immune sensors less strongly than unformulated RNA, but residual impurities (like double-stranded RNA) or high cationic lipid content can still induce inflammation. Careful purification and use of chemical modifications in the RNA (e.g., nucleoside analogs) help reduce unwanted immune activation.

Finally, scale-up and process control are critical for LNP manufacturing. Unlike lab-scale bench mixing, clinical/commercial production uses continuous microfluidic mixing devices. Systems like those from Precision Nanosystems (e.g., NxGen) can process liters of LNP suspension per minute, yielding batches of hundreds of liters.⁷ Continuous flow mixing ensures reproducibility and is easily automated, but it introduces new process parameters. Notably, LNP performance is highly sensitive to mixing conditions: a recent study found that microfluidic vs. manual mixing led to 100-fold differences in in vivo efficacy for some formulations.¹ Each LNP formulation may require unique optimization of flow rates and mixing geometry to balance particle size, encapsulation, and organ tropism.¹ Because mRNA-LNP products are not amenable to terminal sterilization without compromising the payload, commercial production must also be conducted under aseptic GMP conditions.⁸ These conditions impose additional constraints on equipment design and process integration, requiring closed systems, sterile filtration of inputs, validated cleanroom operations, and aseptic fill/finish. In practice, companies often screen formulations at small scale and then transfer to scaled-up microfluidic units, carefully tuning parameters to retain the desired delivery profile.

Emerging Carriers And Targeting Strategies

While LNPs dominate today, alternative carriers are under development to overcome LNP limitations and enable new applications.

Exosomes and extracellular vesicles (EVs) are nanosized vesicles naturally secreted by cells that carry RNA and proteins between cells.⁹ Because they originate from the body’s own cells, exosomes are inherently biocompatible and can carry complex cargo (mRNA, siRNA, proteins). Importantly, some engineered exosomes can cross the blood–brain barrier, offering a potential route for CNS-targeted therapies.⁹ Therapeutic exosomes are typically produced by culturing donor cells (e.g., HEK293 cells or mesenchymal stem cells), which may be engineered to overexpress or selectively package specific RNA cargo. Vesicles are then isolated from conditioned media using methods such as differential ultracentrifugation, size-exclusion chromatography, or tangential flow filtration, the latter offering greater scalability for clinical production. However, because EVs are generated in living cells rather than assembled synthetically, their manufacturing requires controlled cell expansion, upstream bioprocess optimization, and complex downstream purification, creating challenges in scalability, batch-to-batch reproducibility, and GMP compliance compared to fully synthetic nanoparticle systems.

In fact, biotechs are exploring exosome-based RNA therapies: recent industry analyses estimate that more than 70 companies are developing exosome therapeutics, with over 80 pipeline candidates in preclinical or clinical stages.⁹ For example, Aruna Bio’s AB126, derived from neural stem cell exosomes, is entering trials for acute ischemic stroke, and Capricor Therapeutics is developing engineered exosomes as a vaccine and RNA delivery platform.⁹ Despite this momentum, exosome-based approaches face substantial translational hurdles. Standardized large-scale production requires expansion of source cells under controlled conditions and robust downstream purification to ensure identity, potency, and batch consistency.¹⁰ Efficient and reproducible cargo loading remains a key challenge, and regulatory frameworks for complex, cell-derived vesicles are still evolving.¹⁰ Accordingly, scalable isolation strategies such as tangential flow filtration and emerging affinity-based capture methods remain active areas of process innovation.

Polymeric nanoparticles are another class of carriers. Polymers like PLGA, poly(beta-amino ester)s, or biodegradable polycations (e.g., chitosan, polylysine) can form nanoparticles that condense RNA into protective complexes. Polymer carriers offer tunability, allowing chemists to adjust polymer length, branching, and degradability to control release and toxicity. Hybrid approaches are also common: polymer-lipid hybrids or LNPs with polymer shells combine strengths of both materials. The downside is that synthetic polymers may be more complex to manufacture reproducibly, and some (e.g., PEI) can be cytotoxic if not carefully optimized. Research is ongoing into “smart” polymers that respond to pH or enzymes for controlled release.

Peptide- and protein-based carriers include cell-penetrating peptides (CPPs) and engineered proteins that bind RNA. These are often used for targeted delivery: for example, fusing an RNA-binding domain to a targeting ligand (like an antibody fragment) can shuttle RNA to specific cells. Protamine, an FDA-approved polycationic peptide, is already used in some mRNA vaccine formulations to help complex the RNA. Such biomolecule-based systems generally offer excellent biocompatibility but can be harder to formulate as discrete nanoparticles. As a result, many peptide or aptamer systems are still in early development compared to LNPs.

Overall, the industry is pursuing many creative strategies: from incorporating targeting ligands (antibodies, aptamers, sugars) on carriers that home to specific tissues to engineering novel lipid chemistries for targeted tropism^2,11 to using guided nanocarriers (magnetic or ultrasound-responsive particles) to direct RNA to specific sites, these innovations aim to move beyond the liver-centric delivery that current LNPs and GalNAc-conjugates provide.

Key Challenges And Industry Responses

RNA delivery technologies must overcome several practical challenges before broad commercialization. Stability and storage are major concerns. Unformulated RNA is extremely labile, so formulations are usually stored frozen. The COVID-19 mRNA vaccines, for example, required deep-cold supply chains (minus 20 degrees C to minus 80 degrees C). Companies are now developing lyophilized (freeze-dried) LNP formats and incorporating cryoprotectants (sugars like trehalose) to enable fridge- or room-temperature storage. Recent studies report that optimized lyophilized LNPs can remain stable for weeks at ambient temperature, which could greatly ease distribution. In parallel, emerging work shows that incorporating polymeric excipients into dried LNP matrices can further enhance thermostability by reducing RNA hydrolysis and lipid degradation at elevated temperatures,¹² and broader analyses indicate that ionizable lipid chemistry and lyoprotectants strongly influence thermal robustness. Complementary research on nanostructured lipid carriers (NLCs) demonstrates that lyophilized RNA–NLC systems can maintain in vivo activity after prolonged room-temperature storage.¹³ Implementing robust stability protocols (accelerated testing at varying temperatures and humidity) is a best practice in development.

Targeted delivery beyond the liver remains a critical hurdle. Market analyses note that although LNPs and GalNAc conjugates are “highly innovative,” their extrahepatic delivery has been inconsistent.⁴ Most systemic LNPs naturally accumulate in the liver via ApoE recognition. To reach other organs (lung, muscle, tumor, brain), carriers must evade liver clearance and selectively bind new targets. Strategies include adding ligands (e.g., RGD peptides for tumors, antibodies for cell markers) to LNPs or using cell-derived vesicles that carry inherent homing proteins. Each targeting approach must be validated for off-target effects and immunogenicity. Industry practice is to conduct early biodistribution studies (often in animal models) to confirm that engineered carriers reach the intended cells at sufficient levels.

Manufacturing complexity is also a constraint. As one market report observes, “manufacturing complexity is expected to hinder scalability” of next-generation RNA therapeutics.⁴ LNP production involves handling flammable solvents (ethanol), precise fluidics, and tight process control. Small changes in process — lipid purity, mixing rate, solvent ratio) — can lead to variability in particle size or loading. These technical sensitivities are compounded by the requirement for aseptic GMP manufacturing, which necessitates closed systems, sterile filtration of raw materials, and validated cleanroom and fill/finish operations. To manage variability, companies adopt quality by design (QbD) principles and analytical controls: for example, tracking size distribution (by DLS), encapsulation efficiency (via RiboGreen assays), lipid composition (via HPLC), morphology (via electron microscopy), and batch sterility. Analytical methods like HPLC for lipid composition and electron microscopy for morphology are routinely applied. Many developers partner with specialized CDMOs that have GMP-grade lipid libraries and high-throughput mixing equipment.⁷ Indeed, over 50 contract service providers now offer LNP formulation and manufacturing services,⁷ reflecting the modality’s maturation.

Finally, regulatory and quality requirements influence development. Each new delivery technology must meet safety and reproducibility standards. The FDA now expects comprehensive characterization of nanoparticle products, including impurity profiling (e.g., oxidized lipids, truncated RNAs) and immunotoxicity assessments. Best practices include documenting comparability between process scales (e.g., lab batch vs. pilot GMP batch) and establishing stability-indicating assays (to detect subtle changes over time). Early dialogue with regulators (through pre-IND meetings) can clarify acceptable specifications for novel carriers.

Overall, the RNA delivery field is rapidly evolving: new lipid chemistries, machine learning for lipid design, and completely novel carriers appear regularly. Close collaboration between formulation scientists, process engineers, and immunologists will be essential to translate these innovations into globally accessible therapeutics.

*Corresponding authors. Email: jaklenec@mit.edu (A.J.)

References:

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Competing Interests

From FY 2020 to the present, A.J. receives licensing fees (to patents on which she was an inventor) from, invested in, consults (or was on Scientific Advisory Boards or Boards of Directors) for, lectured (and received a fee), or conducts sponsored research at MIT for which she was not paid for the following entities: The Estée Lauder Companies; Moderna Therapeutics; OmniPulse Biosciences; Particles for Humanity; SiO2 Materials Science; VitaKey. 

About The Experts

Chloe Tan is a Chemical-Biological Engineering student at Massachusetts Institute of Technology (MIT) focused on applying engineering principles to complex biological systems. Her interests center on RNA biology and therapeutic innovation, particularly how advances in RNA technologies are redefining drug development and expanding the boundaries of precision medicine. Through both research and writing, she examines the mechanistic and design questions that determine whether an RNA discovery remains conceptual or becomes clinically actionable.

Lauren Levy is a biological engineering student at MIT working at the intersection of RNA delivery, biomaterials, and translational medicine. Her research focuses on how delivery platform design shapes the real-world performance of RNA therapeutics. Through work on microneedle-based mRNA systems and on-patient medical record technologies, she examines how engineering decisions influence scalability, accessibility, and clinical reliability. More broadly, she is interested in translating mechanistic insight into deployable RNA technologies with the potential to broaden access to high-quality healthcare.

Tina Huang is a postdoctoral fellow at MIT in the laboratories of Ana Jaklenec, Ph.D., and Prof. Robert Langer, where she works on next-generation drug delivery systems for RNA, protein, and vaccine therapeutics. Her research focuses on microfluidic fabrication of programmable particles and vesicles, hybrid polymer–lipid systems, and stimuli-responsive materials for controlled and delayed release of biologics.

Ana Jaklenec, Ph.D., a principal investigator at the David H. Koch Institute for Integrative Cancer Research at MIT, is a leader in the fields of bioengineering and materials science, focused on controlled delivery and stability of therapeutics for global health. She has over 15 years of experience and has supervised more than 50 pre- and post-doctoral students. She is an inventor of several drug delivery technologies that have the potential to enable access to medical care globally. The Jaklenec Lab at the Koch Institute is developing new manufacturing techniques for the design of materials at the nano-and micro-scale for self-boosting vaccines, 3D printed on-demand microneedles, heat-stable polymer-based carriers for oral delivery of micronutrients and probiotics, and long-term drug delivery systems for cancer immunotherapy. She has published over 150 manuscripts, patents, and patent applications and has founded three successful companies: Particles for Humanity, VitaKey, and OmniPulse Biosciences. She is the recipient of the NIH NRSA award and is an elected fellow of the National Academy of Inventors, the American Institute of Medical and Biological Engineering, and the Controlled Release Society.