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Review

Metabolism of mRNA Therapeutics: Insights into Pharmacokinetics and Toxicology

1★, 1, 1, 2, 3, 1, 4★ Corresponding author

  1. 1Department of Pharmaceutics, Vishwakarma University, School of Pharmacy, Pune, Maharashtra, INDIA.
  2. 2Department of Mechanical Engineering, Pandit Deendayal Energy University, Gandhinagar, Gujarat, INDIA.
  3. 3Department of Qualtiy Assurance, Pioneer Pharmacy College, Sayajipura, Vadodara, Gujarat, INDIA.
  4. 4Department of Pharmacy, Sumandeep Vidyapeeth Deemed to be University, Piparia, Waghodia, Vadodara, Gujarat, INDIA.

CORRESPONDENCE

Yogita Shinde

Department of Pharmaceutics, Vishwakarma University, School of Pharmacy, Pune, Maharashtra, INDIA.

galandeyogita@gmail.com

Received: 09-08-2025; Revised: 28-10-2025; Accepted: 14-12-2025.

Volume 16, Issue 2 · pp. 517-522 · 2026 · DOI: 10.5530/ijpi.20260539

ABSTRACT

Messenger RNA (mRNA) therapies are a groundbreaking treatment for various conditions, including infectious diseases, cancer, and genetic disorders. This review examines the processing of mRNA therapeutics in the body, focusing on pharmacokinetics, cellular handling, and potential toxicity. It highlights the importance of delivery methods, immune responses, and mRNA stability for effective treatment. Additionally, it addresses challenges like unintended effects and long-term safety, while proposing strategies to enhance mRNA stability and reduce toxicity. Ultimately, it provides insights into how understanding mRNA metabolism can support the future development of these therapies.

KEYWORDS

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FULL TEXT

INTRODUCTION

mRNA therapies are transforming healthcare by enabling cells to produce disease-fighting proteins. Unlike traditional medications, they allow the body to generate therapeutic proteins useful for vaccines, cancer treatment, and genetic disorders. COVID-19 vaccines use mRNA to trigger immune responses, while in cancer, mRNA targets tumor antigens, and for genetic conditions, it aids in restoring essential proteins. Understanding the absorption, distribution, metabolism, and elimination of mRNA is vital for ensuring its safety and effectiveness (Karikó et al., 2008).

mRNA metabolism: an overview

mRNA consists of key components: the 5′ cap, UTRs, ORF, and poly(A) tail, which affect its stability, translation, and which influence its stability, translation efficiency, and therapeutic use, as illustrated in Figure 1 (Mockey 2006; Henderson, 2021; Grier, 2016). The 5′cap, a modified guanine, prevents degradation and aids translation; improved analogs can enhance capping efficiency to 95%, boosting stability and protein production, as demonstrated in Figure 2. The poly(A) tail, consisting of approximately 100–300 adenine residues, also increases stability and can be optimized for better performance (Holtkamp, 2006; Schlake 2012). UTRs regulate translation and can be tailored for better efficiency and tissue-specific expression. While many mRNAs use α- or β-globin UTRs, further optimization can increase yield (Holtkamp, 2006; Vaidyanathan 2018; Ross and Sullivan, 1985).

Modified nucleosides such as 1-methylpseudouridine and N1-methyladenosine boost mRNA stability and lower immune activation. N1-methyladenosine aids in evading RIG-I, which enhances delivery and diminishes the immune response-crucial for progressing mRNA therapies for infections, cancer, and genetic disorders (Pascolo, 2021; Sahin et al., 2014; Kowalski, 2019).

Degradation pathways

Endogenous enzymes control mRNA stability and degradation. Deadenylases shorten the poly(A) tail to start decay, while decapping enzymes DCP1 and DCP2 remove the 5' cap, resulting in degradation by the exosome complex, represented in Figure 3 (Karikó, 2011). Eukaryotic mRNAs degrade from the 3' to 5' end after the poly(A) tail is removed, which is crucial for mRNA stability and translation (Muhlrad et al., 1994; Muhlrad et al., 1995).

In yeast, blocking 5' to 3' decay causes accumulation of 3' PGK1 mRNA fragments, suggesting a shift to 3' to 5' decay. Similar results in oat phytochrome A mRNA indicate this process is conserved (Higgs and Colbert, 1994). The exact roles of exonucleases and endonucleases remain unclear. Notably, some mRNAs display stability without a poly(A) tail, indicating that other factors contribute to stability (Decker and Parker, 1993; Dujon, 1994).

Pharmacokinetics of mRNA therapeutics

Delivery systems and tissue targeting

mRNA delivery efficiency is greatly influenced by the method used, with Lipid Nanoparticles (LNPs) being the most common. LNPs enter cells via endocytosis, releasing mRNA for protein synthesis. Advanced systems like polymeric and hybrid lipid-polymer nanoparticles enhance targeting, while polymeric carriers show promise for lung treatments in asthma and COPD (Hou, 2021; Riley, 2021).

Modern RNA-loaded Lipid Nanoparticles (LNPs), evolved from 1960s liposomes, consist of cholesterol, ionizable lipids, and stealth lipids. Their core features structural lipids, while neutral phospholipids stabilize the bilayer and cholesterol enhances RNA delivery efficiency (Riley, 2021; Ad, 1965; Briuglia, 2015; Gregoriadis and Davis, 1979; Kirby et al., 1980; Malone et al., 1989).

Stability and protein translation

mRNA stability is crucial for its longevity and protein production, especially in therapies. Innovations like Clean Cap, a 5' cap analog, enhance stability, translation efficiency, and degradation resistance, leading to a longer half-life and increased protein synthesis (Wu, 2024; Rouf, 2022). Modifying UTRs boosts stability, aiding single-dose vaccines like Zika by enhancing immune responses and reducing dosage (Richner, 2017; Hengelbrock, 2023). PERSIST-seq involves introducing mRNA pools into HEK293T cells, separating translating from non-translating mRNAs with sucrose gradients. RT-PCR and Illumina sequencing assess translation efficiency, with polysome-associated mRNAs indicating active translation (Kozak, 1994; Babendure, 2006). Early results showed clear translation differences between ActB mRNA and scrambled UTR controls (Osuna, 2017).

Clearance pathways

Post-translation, mRNA breaks down into nucleotides, regulating gene expression and preventing protein accumulation. These nucleotides can be reused or converted into essential metabolites, with excess eliminated to maintain balance (Hoecke and Roose, 2019; Stephenson and Zamecnik, 1978). Clearance mainly involves the kidneys and liver. The kidneys filter waste and salts into urine, while the liver transforms substances into less harmful or water-soluble forms (Sharma and Watts, 2015; Dammes and Peer, 2020; Holland, 2024). The liver produces proteins, regulates glucose storage, and secretes bile for fat digestion, all vital for drug development (Holland, 2024). Understanding nucleotide metabolism aids drug optimization, improving efficacy and safety by balancing clearance rates to reduce dosing frequency and toxicity, while enhancing stability and bioavailability (Watson and Crick, 1953).

Toxicological implications

Immunogenic responses

Unaltered mRNA activates receptors like TLR3, TLR7, TLR8, and RIG-I, causing inflammation or autoimmune responses. Modified nucleosides, such as 5-methylcytidine and pseudouridine, enhance stability, reduce immune detection, and improve translation (Moles, 2019; Mu and Hur, 2021). Enhanced delivery boosts safety and effectiveness in vaccines, cancer treatments, and genetic therapies (Haque, 2020; Heine et al., 2021). Continuous research focuses on improving carriers and minimizing immune reactions (Corbett, 2020; Maruggi, 2019).

Cumulative toxicity

The use of mRNA treatments, like vaccines and gene therapy, raises concerns about Lipid Nanoparticles (LNPs) accumulating in the liver and spleen, which are crucial for detoxification and immune function (Fang, 2022; Hu, 2020; Janas, 2018; Lv, 2006; Monnery, 2017). LNP accumulation can lead to lasting toxicity. To mitigate this, researchers are creating biodegradable carriers that decompose into safe by products, enhancing safety and delivery (Fischer, 2003; Moghimi, 2005; Kedmi, 2010; Soenen, 2009).

siRNAs can also induce immune responses and toxicity, complicating their clinical use (Dokka, 2000). Pattern Recognition Receptors (PRRs), such as Toll-like Receptors (TLRs), trigger cytokine release, inflammation, and reduced effectiveness (Dokka, 2000). Unmodified siRNAs can cause unintended effects, stress, and cell death, and certain delivery methods may heighten immune activation (Toh, 2009; Horie, 2013).

Potential integration risks

mRNA plays a crucial role in protein synthesis and does not change the host genome, making it suitable for vaccines and treatments (Wolff, 1990). Its stability and translation are influenced by the sequence, cell type, and surrounding environment (Boczkowski, 1996). Since 1990, the use of exogenous mRNA has led to significant advancements in therapeutic and preventive molecular medicine (Hoerr, 2000).

RNA technologies currently facilitate cancer immunotherapy, protein replacement, genome editing, and vaccine development (Hu, 2020). mRNA is responsible for encoding proteins that can fix mutations or enhance immune responses, while siRNA is used to silence detrimental genes associated with diseases (Chavda, 2021). To increase safety, current designs incorporate degradation signals to manage the lifespan of mRNA (Sahin, 2014), striking a balance between effectiveness and risk (Kreiter, 2016). Ongoing studies are focused on improving the performance of mRNA therapies (Zheng, 2000; Vader, 2016).

Emerging strategies to optimize mRNA therapeutics

Advanced delivery systems

Exosomes are natural vesicles that carry biomolecules such as mRNA, providing biocompatibility and targeted gene delivery for cancer with little effect on healthy cells, as depicted in Figure 4 (Sedic, 2018). Dendritic polymers, also known as dendrimers, are branched nanocarriers that can deliver small molecules, proteins, and nucleic acids with great accuracy and absorption. Merging these two systems could improve the effectiveness and safety of therapies, especially in cancer treatment (Li, 2014; Funabiki, 2014).

Bioconjugation technologies

Attaching specific molecules to mRNA delivery systems enhances accuracy by focusing on particular tissues and minimizing systemic side effects. Lipid Nanoparticles (LNPs), which are frequently utilized for mRNA delivery, can be altered with ligands such as galactose to specifically target liver cells through receptor recognition. This approach is particularly beneficial for addressing rare liver-related metabolic disorders by delivering mRNA that encodes functional proteins directly to hepatocytes, thereby restoring function and alleviating symptoms along with off-target effects (Ramírez-Cortés and Ménová, 2025).

Crispr-driven optimization

Attaching specific molecules to mRNA delivery systems enhances the accuracy and effectiveness of mRNA therapeutics by allowing targeted delivery to specific tissues, which helps to minimize side effects from systemic distribution. Lipid Nanoparticles (LNPs) are a prominent delivery method, encapsulating mRNA and facilitating cellular entry. When modified with targeting ligands such as galactose-recognized by liver cell receptors-LNPs improve liver-specific delivery. This is particularly beneficial for treating liver-related metabolic disorders resulting from genetic mutations. The targeted delivery of mRNA that encodes functional proteins can restore liver function, alleviate symptoms, and reduce necessary dosages, thereby minimizing off-target effects (Ramírez-Cortés and Ménová, 2025).

Figure 1: Messenger RNA (mRNA) (Chavda, 2021).
Figure 2: Post-transcriptional capping (left) is a two-step enzymatic process, whereas co-transcriptional capping (right) adds a cap analog during transcription using an enzyme from in vitro transcription (Hu, 2020).
Figure 3: Eukaryotes degrade mRNA to regulate gene expression and control protein production in response to changing conditions (Fischer, 2003).
Figure 4: There are various methods for packing cargo into exosomes (Van Hoecke and Roose, 2019).

CONCLUSION

mRNA therapies mark a significant breakthrough in the treatment of infectious diseases and genetic disorders. Their effectiveness depends on a solid grasp of pharmacokinetics, processing, and possible toxicity. Progress in chemical modifications, delivery methods, and translation efficiency has enhanced stability, minimized immune responses, and allowed for targeted actions. Their applications include vaccines, cancer therapies, and rare genetic disorders, although concerns regarding long-term safety and immune activation remain and immune activation. Emerging tools such as exosome-based delivery and CRISPR systems further expand therapeutic potential. Ongoing collaboration across disciplines is crucial to ensure a balance between safety and effectiveness in future mRNA therapies.

Acknowledgement

The authors would like to express their gratitude to the faculty members and management of Vishwakarma University, School of Pharmacy, Pune, Maharashtra, India for providing all necessary technical supports and motivating us throughout the project.

Funding

This review work is not financially supported by any funding agency.

Conflict of Interest

The authors declare that there is no conflict of interest.

Data Availability

All data supporting the findings of this review are contained within the article itself. No additional datasets were generated or analyzed during the preparation of this manuscript. References to external sources and literature are appropriately cited throughout the text.

Ethics Statement

Not applicable.

REFERENCES

As published

Showing references and in-text citations exactly as published.

  1. Boczkowski, D, Nair, S. K, Snyder, D, & Gilboa, E. (1996). Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo. The Journal of Experimental Medicine, 184(2), 465–472. https://doi.org/10.1084/jem.184.2.465CrossrefGOOGLE SCHOLAR
  2. Briuglia, M, Rotella, C, McFarlane, A, & Lamprou, D. A. (2015). Influence of cholesterol on liposome stability and on in vitro drug release. Drug Delivery and Translational Research, 5(3), 231–242. https://doi.org/10.1007/s13346-015-0220-8CrossrefGOOGLE SCHOLAR
  3. Chavda, V. P, Hossain, M. K, Beladiya, J, & Apostolopoulos, V. (2021). Nucleic acid vaccines for COVID-19: A paradigm shift in the vaccine development arena. Biologics, 1(3), 337–356. https://doi.org/10.3390/biologics1030020CrossrefGOOGLE SCHOLAR
  4. Corbett, K. (2020). S, Edwards, D. K, Leist, S. R, Abiona, O. M, Boyoglu-barnum, S, Gillespie, R. A, Himansu, S, Schäfer, A, Ziwawo, C. T, Dipiazza, A. T, Dinnon, K. H, Elbashir, S. M, Shaw, C. A, Woods, A, Fritch, E. J, Martinez, D. R, Bock, K. W, Minai, M, Nagata, B. M, Graham, B. S. SARS-CoV-2 mRNA Vaccine Design Enabled By Prototype Pathogen Preparedness. Nature, 586(7830), 567–571. https://doi.org/10.1038/s41586-020-2622-0CrossrefGOOGLE SCHOLAR
  5. Decker, C. J, & Parker, R. (1993). A turnover pathway for both stable and unstable MRNAs in yeast: Evidence for a requirement for deadenylation. Genes and Development, 7(8), 1632–1643. https://doi.org/10.1101/gad.7.8.1632CrossrefGOOGLE SCHOLAR
  6. Dokka, S, Toledo, D, Shi, X, Castranova, V, & Rojanasakul, Y. (2000). Oxygen radical-mediated pulmonary toxicity induced by some cationic liposomes. Pharmaceutical Research, 17(5), 521–525. https://doi.org/10.1023/A:1007504613351CrossrefGOOGLE SCHOLAR
  7. Dörrie, J, Schaft, N, Schuler, G, & Schuler-Thurner, B. (2020). Therapeutic cancer vaccination with ex vivo RNA-transfected dendritic cells-An update. Pharmaceutics, 12(2), Article, 92. https://doi.org/10.3390/pharmaceutics12020092CrossrefGOOGLE SCHOLAR
  8. Dujon, B, Alexandraki, D, André, B, Ansorge, W, Baladron, V, & Ballesta, J. (1994). P. G, Banrevi, A, Bolle, P. A, Bolotin-fukuhara, M, Bossier, P, Bou, G, Boyer, J, Bultrago, M. J, Cheret, G, Colleaux, L, Dalgnan-fornler, B, Del Rey, F, Dlon, C, Domdey, H, Mewes, H. W. Complete DNA Sequence of Yeast Chromosome XI. Nature, 369(6479), 371–378. https://doi.org/10.1038/369371a0CrossrefGOOGLE SCHOLAR
  9. Fang, E, Liu, X, Li, M, Zhang, Z, Song, L, Zhu, B, Wu, X, Liu, J, Zhao, D, & Li, Y. (2022). Advances in COVID-19 MRNA vaccine development. Signal Transduction and Targeted Therapy, 7(1), Article, 94. https://doi.org/10.1038/s41392-022-00950-yCrossrefGOOGLE SCHOLAR
  10. Fischer, D, Li, Y, Ahlemeyer, B, Krieglstein, J, & Kissel, T. (2003). In vitro cytotoxicity testing of polycations: Influence of polymer structure on cell viability and hemolysis. Biomaterials, 24(7), 1121–1131. )00445-3. https://doi.org/10.1016/S0142-9612(02CrossrefGOOGLE SCHOLAR
  11. Funabiki, M, Kato, H, Miyachi, Y, Toki, H, Motegi, H, Inoue, M, Minowa, O, Yoshida, A, Deguchi, K, Sato, H, Ito, S, Shiroishi, T, Takeyasu, K, Noda, T, & Fujita, T. (2014). (2014). Autoimmune Disorders Associated With Gain of Function of the Intracellular Sensor MDA5. Immunity, 40(2), 199–212. https://doi.org/10.1016/j.immuni.2013.12.014CrossrefGOOGLE SCHOLAR
  12. Gregoriadis, G, & Davis, C. (1979). Stability of liposomes invivo and invitro is promoted by their cholesterol content and the presence of blood cells. Biochemical and Biophysical Research Communications, 89(4), 1287–1293. )92148-X. https://doi.org/10.1016/0006-291X(79CrossrefGOOGLE SCHOLAR
  13. Grier, A. E, Burleigh, S, Sahni, J, Clough, C. A, Cardot, V, Choe, D. C, Krutein, M. C, Rawlings, D. J, Jensen, M. C, Scharenberg, A. M, & Jacoby, K. (2016). PEVL: A linear plasmid for generating MRNA IVT templates with extended encoded poly(A) sequences. Molecular Therapy. Nucleic Acids, 5(4), e306. https://doi.org/10.1038/mtna.2016.21CrossrefGOOGLE SCHOLAR
  14. Haque, A. A, Weinmann, P, Biswas, S, Handgretinger, R, Mezger, M, Kormann, M. S. D, & Antony, J. S. (2020). RNA ImmunoGenic Assay: Simple method for detecting immunogenicity of in vitro transcribed MRNA. Advances in Cell and Gene Therapy, 3(2), e79. https://doi.org/10.1002/acg2.79CrossrefGOOGLE SCHOLAR
  15. Heine, A, Juranek, S, & Brossart, P. (2021). Clinical and immunological effects of MRNA vaccines in malignant diseases. Molecular Cancer, 20(1), Article, 52. https://doi.org/10.1186/s12943-021-01339-1CrossrefGOOGLE SCHOLAR
  16. Henderson, J. M, Ujita, A, Hill, E, Yousif-Rosales, S, Smith, C, Ko, N, McReynolds, T, Cabral, C. R, Escamilla-Powers, J. R, & Houston, M. E. (2021). Cap 1 messenger RNA synthesis with co‐transcriptional cleancap® analog by in vitro transcription. Current Protocols, 1(2), e39. https://doi.org/10.1002/cpz1.39CrossrefGOOGLE SCHOLAR
  17. Hengelbrock, A, Schmidt, A, Helgers, H, Vetter, F. L, & Strube, J. (2023). Scalable MRNA machine for regulatory approval of variable scale between 1000 clinical doses to 10 million manufacturing scale doses. Processes, 11(3), 745. https://doi.org/10.3390/pr11030745CrossrefGOOGLE SCHOLAR
  18. Higgs, D. C, & Colbert, J. T. (1994). Oat phytochrome A MRNA degradation appears to occur via two distinct pathways. The Plant Cell, 6(7), 1007–1019. https://doi.org/10.1105/tpc.6.7.1007CrossrefGOOGLE SCHOLAR
  19. Hoecke, V, L, & Roose, K. (2019). How MRNA therapeutics are entering the monoclonal antibody field. Journal of Translational Medicine, 17(1), Article, 54. https://doi.org/10.1186/s12967-019-1804-8CrossrefGOOGLE SCHOLAR
  20. Hoerr, I, Obst, R, Rammensee, H. G, & Jung, G. (2000). In vivo application of RNA leads to induction of specific cytotoxic T lymphocytes and antibodies. European Journal of Immunology, 30(1), 1–7. )30:13.0.CO;2-#. https://doi.org/10.1002/1521-4141(200001CrossrefGOOGLE SCHOLAR
  21. Holland, R, Lam, K, Jeng, S, McClintock, K, Palmer, L, Schreiner, P, Wood, M, Zhao, W, & Heyes, J. (2024). Silicon ether ionizable lipids enable potent MRNA lipid nanoparticles with rapid tissue clearance. ACS Nano, 18(15), 10374–10387. https://doi.org/10.1021/acsnano.3c09028CrossrefGOOGLE SCHOLAR
  22. Holtkamp, S, Kreiter, S, Selmi, A, Simon, P, Koslowski, M, Huber, C, Türeci, O, & Sahin, U. (2006). Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells. Blood, 108(13), 4009–4017. https://doi.org/10.1182/blood-2006-04-015024CrossrefGOOGLE SCHOLAR
  23. Horie, M, Kobayashi, Y, Suzuki, Y, & Tomonaga, K. (2013). Comprehensive analysis of endogenous bornavirus-like elements in eukaryote genomes. Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences, 368(1626), Article 20120499. https://doi.org/10.1098/rstb.2012.0499CrossrefGOOGLE SCHOLAR
  24. Hou, X, Zaks, T, Langer, R, & Dong, Y. (2021). Lipid nanoparticles for MRNA delivery. Nature Reviews. Materials, 6(12), 1078–1094. https://doi.org/10.1038/s41578-021-00358-0CrossrefGOOGLE SCHOLAR
  25. Hu, B, Zhong, L, Weng, Y, Peng, L, Huang, Y, Zhao, Y, & Liang, X. (2020). Therapeutic SiRNA: State of the art. Signal Transduction and Targeted Therapy, 5(1), Article, 101. https://doi.org/10.1038/s41392-020-0207-xCrossrefGOOGLE SCHOLAR
  26. Janas, M. (2018). M, Schlegel, M. K, Harbison, C. E, Yilmaz, V. O, Jiang, Y, Parmar, R, Zlatev, I, Castoreno, A, Xu, H, Shulga-morskaya, S, Rajeev, K. G, Manoharan, M, Keirstead, N. D, Maier, M. A, Jadhav, V. Selection of GalNAc-conjugated siRNAs With Limited Off-target-driven Rat Hepatotoxicity. Nature Communications, 9(1), Article, 723. https://doi.org/10.1038/s41467-018-02989-4CrossrefGOOGLE SCHOLAR
  27. Karikó, K, Muramatsu, H, Ludwig, J, & Weissman, D. (2011). Generating the optimal MRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding MRNA. Nucleic Acids Research, 39(21), e142. -e142. https://doi.org/10.1093/nar/gkr695CrossrefGOOGLE SCHOLAR
  28. Kedmi, R, Ben-Arie, N, & Peer, D. (2010). The systemic toxicity of positively charged lipid nanoparticles and the role of toll-like receptor 4 in immune activation. Biomaterials, 31(26), 6867–6875. https://doi.org/10.1016/j.biomaterials.2010.05.027CrossrefGOOGLE SCHOLAR
  29. Kirby, C, Clarke, J, & Gregoriadis, G. (1980). Effect of the cholesterol content of small unilamellar liposomes on their stability in vivo and in vitro. The Biochemical Journal, 186(2), 591–598. https://doi.org/10.1042/bj1860591CrossrefGOOGLE SCHOLAR
  30. Kowalski, P. S, Rudra, A, Miao, L, & Anderson, D. G. (2019). Delivering the messenger: Advances in technologies for therapeutic MRNA delivery. Molecular Therapy, 27(4), 710–728. https://doi.org/10.1016/j.ymthe.2019.02.012CrossrefGOOGLE SCHOLAR
  31. Kozak, M. (1994). Features in the 5′ non-coding sequences of rabbit α and β-globin MRNAs that affect translational efficiency. Journal of Molecular Biology, 235(1), 95–110. )80019-1. https://doi.org/10.1016/S0022-2836(05CrossrefGOOGLE SCHOLAR
  32. Kreiter, S, Diken, M, Pascolo, S, Nair, S. K, Thielemans, K. M, & Geall, A. (2016). RNA vaccination therapy: Advances in an emerging field. Journal of Immunology Research, Article 9703914. https://doi.org/10.1155/2016/9703914CrossrefGOOGLE SCHOLAR
  33. Li, F, Nellåker, C, Sabunciyan, S, Yolken, R. H, Jones-Brando, L, Johansson, A, Owe-Larsson, B, & Karlsson, H. (2014). Transcriptional derepression of the ERVWE1 locus following influenza A virus infection. Journal of Virology, 88(8), 4328–4337. https://doi.org/10.1128/JVI.03628-13CrossrefGOOGLE SCHOLAR
  34. Lindel, F, Dodt, C. R, Weidner, N, Noll, M, Bergemann, F, Behrendt, R, Fischer, S, Dietrich, J, Cartellieri, M, Hamann, M. V, & Lindemann, D. (2019). TraFo-CRISPR: Enhanced genome engineering by transient foamy virus vector-mediated delivery of CRISPR/Cas9 components. Molecular Therapy. Nucleic Acids, 18, 708–726. https://doi.org/10.1016/j.omtn.2019.10.006CrossrefGOOGLE SCHOLAR
  35. Litzinger, D. C, Brown, J. M, Wala, I, Kaufman, S. A, Van, G. Y, Farrell, C. L, & Collins, D. (1996). Fate of cationic liposomes and their complex with oligonucleotive in vivo. Biochimica et Biophysica Acta, 1281(2), 139–149. )00268-5. https://doi.org/10.1016/0005-2736(95CrossrefGOOGLE SCHOLAR
  36. Lv, H, Zhang, S, Wang, B, Cui, S, & Yan, J. (2006). Toxicity of cationic lipids and cationic polymers in gene delivery. Journal of Controlled Release, 114(1), 100–109. https://doi.org/10.1016/j.jconrel.2006.04.014CrossrefGOOGLE SCHOLAR
  37. Malone, R. W, Felgner, P. L, & Verma, I. M. (1989). Proceedings of the national academy of sciences of the united states of America, 86(16), 6077-6081. https://doi.org/10.1073/pnas.86.16.6077CrossrefGOOGLE SCHOLAR
  38. Manfredonia, I, & Incarnato, D. (2021). Structure and regulation of coronavirus genomes: State-of-the-art and novel insights from SARS-CoV-2 studies. Biochemical Society Transactions, 49(1), 341–352. https://doi.org/10.1042/BST20200670CrossrefGOOGLE SCHOLAR
  39. Marasca, F, Sinha, S, Vadalà, R, Polimeni, B, Ranzani, V, & Paraboschi, E. (2022). M, Burattin, F. V, Ghilotti, M, Crosti, M, Negri, M. L, Campagnoli, S, Notarbartolo, S, Sartore-bianchi, A, Siena, S, Prati, D, Montini, G, Viale, G, Torre, O, Harari, S, Bodega, B. LINE1 Are Spliced in Non-canonical Transcript Variants to Regulate T Cell Quiescence and Exhaustion. Nature Genetics, 54(2), 180–193. LINE1. https://doi.org/10.1038/s41588-021-00989-7CrossrefGOOGLE SCHOLAR
  40. Martier, R. M. (2016). Therapeutic RNAi-based gene therapy for neurodegenerative disorders: Slowing. Neuropathology, 30, 103–112.GOOGLE SCHOLAR
  41. Maruggi, G, Zhang, C, Li, J, Ulmer, J. B, & Yu, D. (2019). MRNA as a transformative technology for vaccine development to control infectious diseases. Molecular Therapy, 27(4), 757–772. https://doi.org/10.1016/j.ymthe.2019.01.020CrossrefGOOGLE SCHOLAR
  42. Moghimi, S. M, Symonds, P, Murray, J. C, Hunter, A. C, Debska, G, & Szewczyk, A. (2005). A two-stage poly(ethylenimine)-mediated cytotoxicity: Implications for gene transfer/therapy. Molecular Therapy, 11(6), 990–995. https://doi.org/10.1016/j.ymthe.2005.02.010CrossrefGOOGLE SCHOLAR
  43. Moles, E, Kavallaris, M, & Fernàndez-Busquets, X. (2019). Modeling the distribution of diprotic basic drugs in liposomal systems: Perspectives on malaria nanotherapy. Frontiers in Pharmacology, 10, Article, 1064. https://doi.org/10.3389/fphar.2019.01064CrossrefGOOGLE SCHOLAR
  44. Monnery, B. D, Wright, M, Cavill, R, Hoogenboom, R, Shaunak, S, Steinke, J. H. G, & Thanou, M. (2017). Cytotoxicity of polycations: Relationship of molecular weight and the hydrolytic theory of the mechanism of toxicity. International Journal of Pharmaceutics, 521(1–2), 249–258. https://doi.org/10.1016/j.ijpharm.2017.02.048CrossrefGOOGLE SCHOLAR
  45. Mu, X, & Hur, S. (2021). Immunogenicity of in vitro-transcribed RNA. Accounts of Chemical Research, 54(21), 4012–4023. https://doi.org/10.1021/acs.accounts.1c00521CrossrefGOOGLE SCHOLAR
  46. Muhlrad, D, Decker, C. J, & Parker, R. (1994). Deadenylation of the unstable MRNA encoded by the yeast MFA2 gene leads to decapping followed by 5′→3′ digestion of the transcript. Genes and Development, 8(7), 855–866. https://doi.org/10.1101/gad.8.7.855CrossrefGOOGLE SCHOLAR
  47. Muhlrad, D, Decker, C. J, & Parker, R. (1995). Turnover mechanisms of the stable yeast PGK1 MRNA. Molecular and Cellular Biology, 15(4), 2145–2156. https://doi.org/10.1128/MCB.15.4.2145CrossrefGOOGLE SCHOLAR
  48. Osuna, B. A, Howard, C. J, Kc, S, Frost, A, & Weinberg, D. E. (2017). In vitro analysis of RQC activities provides insights into the mechanism and function of CAT tailing. Elife, 6, Article E27949. https://doi.org/10.7554/eLife.27949CrossrefGOOGLE SCHOLAR
  49. Pardi, N, & Krammer, F. (2024). MRNA vaccines for infectious diseases-Advances, challenges and opportunities. Nature Reviews. Drug Discovery, 23(11), 838–861. https://doi.org/10.1038/s41573-024-01042-yCrossrefGOOGLE SCHOLAR
  50. Pardi, N, Tuyishime, S, Muramatsu, H, Kariko, K, Mui, B. L, Tam, Y. K, Madden, T. D, Hope, M. J, & Weissman, D. (2015). Expression kinetics of nucleoside-modified MRNA delivered in lipid nanoparticles to mice by various routes. Journal of Controlled Release, 217, 345–351. https://doi.org/10.1016/j.jconrel.2015.08.007CrossrefGOOGLE SCHOLAR
  51. Pascolo, S. (2021). Vaccines against COVID-19: Priority to MRNA-based formulations. Cells, 10(10), Article, 2716. https://doi.org/10.3390/cells10102716CrossrefGOOGLE SCHOLAR
  52. Ramírez-Cortés, F, & Ménová, P. (2025). Hepatocyte targeting via the asialoglycoprotein receptor. RSC Medicinal Chemistry, 16(2), 525–544. https://doi.org/10.1039/D4MD00652FCrossrefGOOGLE SCHOLAR
  53. Richner, J. (2017). M, Himansu, S, Dowd, K. A, Butler, S. L, Salazar, V, Fox, J. M, Julander, J. G, Tang, W. W, Shresta, S, Pierson, T. C, Ciaramella, G, Diamond, M. S. Modified mRNA Vaccines Protect Against Zika Virus Infection. Cell, 168(6), 1114–1125. e10. https://doi.org/10.1016/j.cell.2017.02.017CrossrefGOOGLE SCHOLAR
  54. Riley, R. (2021). S, Kashyap, M. V, Billingsley, M. M, White, B, Alameh, M, Bose, S. K, Zoltick, P. W, Li, H, Zhang, R, Cheng, A. Y, Weissman, D, Peranteau, W. H, Mitchell, M. J. Ionizable Lipid Nanoparticles for in Utero mRNA Delivery. Science Advances, 7(3), Article Eaba1028. https://doi.org/10.1126/sciadv.aba1028CrossrefGOOGLE SCHOLAR
  55. Rouf, N. Z, Biswas, S, Tarannum, N, Oishee, L. M, & Muna, M. M. (2022). Demystifying MRNA vaccines: An emerging platform at the forefront of cryptic diseases. RNA Biology, 19(1), 386–410. https://doi.org/10.1080/15476286.2022.2055923CrossrefGOOGLE SCHOLAR
  56. Sahin, U, Karikó, K, Türeci, & Ö (2014). MRNA-based therapeutics—Developing a new class of drugs. Nature Reviews. Drug Discovery, 13(10), 759–780. https://doi.org/10.1038/nrd4278CrossrefGOOGLE SCHOLAR
  57. Sedic, M, Senn, J. J, Lynn, A, Laska, M, Smith, M, Platz, S. J, Bolen, J, Hoge, S, Bulychev, A, Jacquinet, E, Bartlett, V, & Smith, P. F. (2018). Safety evaluation of lipid nanoparticle-formulated modified MRNA in the Sprague-Dawley rat and cynomolgus monkey. Veterinary Pathology, 55(2), 341–354. https://doi.org/10.1177/0300985817738095CrossrefGOOGLE SCHOLAR
  58. Sercombe, L, Veerati, T, Moheimani, F, Wu, S. Y, Sood, A. K, & Hua, S. (2015). Advances and challenges of liposome assisted drug delivery. Frontiers in Pharmacology, 6, Article, 286. https://doi.org/10.3389/fphar.2015.00286CrossrefGOOGLE SCHOLAR
  59. Sharma, V. K, & Watts, J. K. (2015). Oligonucleotide therapeutics: Chemistry, delivery and clinical progress. Future Medicinal Chemistry, 7(16), 2221–2242. https://doi.org/10.4155/fmc.15.144CrossrefGOOGLE SCHOLAR
  60. Soenen, S. J. H, Brisson, A. R, Cuyper, D, & M (2009). Addressing the problem of cationic lipid-mediated toxicity: The magnetoliposome model. Biomaterials, 30(22), 3691–3701. https://doi.org/10.1016/j.biomaterials.2009.03.040CrossrefGOOGLE SCHOLAR
  61. Stephenson, M. L, & Zamecnik, P. C. (1978). Proceedings of the national academy of sciences of the united states of America, 75(1), 285-288. https://doi.org/10.1073/pnas.75.1.285CrossrefGOOGLE SCHOLAR
  62. Toh, Y, Lim, T. C, Tai, D, Xiao, G, Noort, V, D, & Yu, H. (2009). A microfluidic 3D hepatocyte chip for drug toxicity testing. Lab on a Chip, 9(14), 2026–2035. https://doi.org/10.1039/b900912dCrossrefGOOGLE SCHOLAR
  63. Tuller, T, & Zur, H. (2015). Multiple roles of the coding sequence 5′ end in gene expression regulation. Nucleic Acids Research, 43(1), 13–28. https://doi.org/10.1093/nar/gku1313CrossrefGOOGLE SCHOLAR
  64. Vader, P, Mol, E. A, Pasterkamp, G, & Schiffelers, R. M. (2016). Extracellular vesicles for drug delivery. Advanced Drug Delivery Reviews, 106(A), 148–156. https://doi.org/10.1016/j.addr.2016.02.006CrossrefGOOGLE SCHOLAR
  65. Warrier, D. U, Desai, H. R, Singh, K. H, & Shinde, U. A. (2021). Solid lipid nanoparticle-based drug delivery for lung cancer. In Handbook of Lung Targeted Drug Delivery Systems, 303-, 323. https://doi.org/10.1201/9781003046547-21CrossrefGOOGLE SCHOLAR
  66. Watson, J. D, & Crick, F. H. (1953). A structure for ADN. Nature, 171(4356), 737–738. https://doi.org/10.1038/171737a0CrossrefGOOGLE SCHOLAR
  67. Wolff, J. A, Malone, R. W, Williams, P, Chong, W, Acsadi, G, Jani, A, & Felgner, P. L. (1990). Direct gene transfer into mouse muscle in vivo. Science, 247(4949 Pt 1), 1465–1468. https://doi.org/10.1126/science.1690918CrossrefGOOGLE SCHOLAR
  68. Wu, L, Li, X, Qian, X, Wang, S, Liu, J, & Yan, J. (2024). Lipid nanoparticle (LNP) delivery carrier-assisted targeted controlled release MRNA vaccines in tumor immunity. Vaccines, 12(2), Article, 186. https://doi.org/10.3390/vaccines12020186CrossrefGOOGLE SCHOLAR
  69. Zheng, C, Baum, B. J, Iadarola, M. J, & O’Connell, B. C. (2000). Genomic integration and gene expression by a modified adenoviral vector. Nature Biotechnology, 18(2), 176–180. https://doi.org/10.1038/72628CrossrefGOOGLE SCHOLAR

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Shinde, Y., Sharma, T. P., Shaha, A., Chaudhari, R., Rajeghorpade, R., Talele, D., & Talele, C. (2026). Metabolism of mRNA Therapeutics: Insights into Pharmacokinetics and Toxicology. International Journal of Pharmaceutical Investigation, 16(2), 517-522. https://doi.org/10.5530/ijpi.20260539

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