This in-depth PowerPoint presentation explores the rapidly advancing field of mRNA vaccine technology, extending well beyond COVID-19. Created by a postgraduate in microbiology, this academic resource outlines the biological foundation, mechanisms of action, types of mRNA vaccines, and advanced delivery systems like lipid nanoparticles, peptides, polymers, and exosomes. The presentation also highlights a wide array of emerging clinical applications, including: Infectious disease vaccines (e.g., Zika, TB, rabies) Cancer immunotherapy using neoantigen-based mRNA platforms Autoimmune & rare genetic disorder therapies Protein replacement therapy Regenerative medicine and neurodegenerative disease treatments Gene editing and CRISPR-assisted therapeutics Future roles in pandemic preparedness and biodefense Special emphasis is placed on self-amplifying (saRNA) and trans-amplifying (taRNA) vaccines, along with Indian mRNA vaccine developments like GEMCOVAC®. Suitable for medical students, microbiologists, biotech professionals, and public health educators, this slide deck serves as a powerful learning and teaching tool in molecular vaccinology and therapeutic biotechnology.

1. mRNA vaccine technology &
its applications beyond COVID-19
Dr Adarsh S
3rd Year PG student
Dept. of Microbiology
ESIC MC & PGIMSR
Rajajinagar, Bangalore

2. References
Al Fayez, N.; Nassar, M.S.; Alshehri, A.A.; Alnefaie, M.K.; Almughem, F.A.; Alshehri, B.Y.; Alawad, A.O.; Tawfik,
E.A. Recent Advancement in mRNA Vaccine Development and Applications. Pharmaceutics 2023, 15, 1972.
Wang YS, Kumari M, Chen GH, Hong MH, Yuan JP, Tsai JL, Wu HC. mRNA-based vaccines and therapeutics: an
in-depth survey of current and upcoming clinical applications. J Biomed Sci. 2023 Oct 7;30(1):84.
Chandra S, Wilson JC, Good D, Wei MQ. mRNA vaccines: a new era in vaccine development. Oncol Res. 2024 Sep
18;32(10):1543-1564
Parhiz H, Atochina-Vasserman EN, Weissman D. mRNA-based therapeutics: looking beyond COVID-19 vaccines.
Lancet. 2024 Mar 23;403(10432):1192-1204

3. Chen B, Yang Y, Wang X, Yang W, Lu Y, Wang D, Zhuo E, Tang Y, Su J, Tang G, Shao S, Gu K. mRNA
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Ankrah PK, Ilesanmi A, Akinyemi AO, Lasehinde V, Adurosakin OE, Ajayi OH. Clinical Analysis and
Applications of mRNA Vaccines in Infectious Diseases and Cancer Treatment. Cureus. 2023 Oct
2;15(10):e46354.
Pardi N, Hogan MJ, Porter FW, Weissman D. mRNA vaccines - a new era in vaccinology. Nat Rev Drug
Discov. 2018 Apr;17(4):261-279.
You H, Jones MK, Gordon CA, Arganda AE, Cai P, Al-Wassiti H, Pouton CW, McManus DP. The mRNA
Vaccine Technology Era and the Future Control of Parasitic Infections. Clin Microbiol Rev. 2023 Mar
23;36(1)

4. Layout
 Central Dogma of Molecular Biology
 mRNA vaccine
 Advantages of mRNA Vaccines
 Disadvantages of mRNA Vaccines
 Types of mRNA Vaccines
 Mechanism of Action of mRNA Vaccines
 The mRNA Vaccine Delivery Systems
Polymeric Nanoparticles
Peptides and Proteins Nanoparticles
Protamine Nanoparticles
Lipid Nanoparticles (LNPs)
Exosome-based systems
 The mRNA Vaccine Applications
Infectious Disease Vaccines
Autoimmune & Rare Disease Therapies
Cancer Immunotherapy
Protein Replacement Therapy
Regenerative Medicine
Neurodegenerative Disease Therapies
Pandemic Preparedness & Biodefense
Gene Editing & CRISPR Applications

5. Purines
Pyramidines

6. Central Dogma of Molecular Biology

10. An mRNA vaccine is a type of vaccine that uses messenger RNA (mRNA) to instruct cells to produce a
specific antigenic protein, triggering an immune response without using a live pathogen.
Open Reading Frame

11. Katalin Karikó and Drew Weissman

12. Mechanism of Action of mRNA Vaccines

14. A conventional vaccine uses whole or parts of a pathogen (virus or bacteria) to stimulate the immune system and
provide immunity against infectious diseases.
Types
1.Live Attenuated Vaccines Use a weakened
form of the pathogen that can replicate but
does not cause disease.
MMR (Measles, Mumps, Rubella), BCG
(Tuberculosis), Oral Polio Vaccine (OPV).
2.Inactivated (Killed) Vaccines – Contain
whole pathogens that have been killed or
inactivated.
Inactivated Polio Vaccine (IPV), Hepatitis A
vaccine.
3. Subunit, Recombinant, or Conjugate Vaccines – Contain only
specific parts (antigens) of a pathogen.
Hepatitis B vaccine, HPV vaccine, Pneumococcal vaccine.
4. Toxoid Vaccines – Use inactivated bacterial toxins to generate
immunity. Tetanus and Diphtheria vaccines.

15. Advantages of mRNA Vaccines Over Conventional Vaccines
1. Faster Development and Deployment
2. No Need for Adjuvants
3. Safer for Immunocompromised Individuals
4. Stronger and More Balanced Immune Response
5. No Anti-Vector Immunity Issues
6. More Precise Antigen Expression
7. Reduced Risk of Allergic Reactions and Egg-Based
Contaminants
8. No Need for a Live Pathogen
9. No Risk of Genomic Integration
10. Easier Modification and Adaptability
11. Scalable and Cost-Effective Production
Disadvantages of mRNA Vaccines
1. Cold Storage and Stability Issues
2. Short mRNA Half-Life
3. Potential for Stronger Side Effects
4. Limited Long-Term Data
5. Lower Stability Compared to Conventional
Vaccines
6. Potential for Unintended Immune Reactions
7. Risk of Autoimmune Reactions
8. Manufacturing and Scalability Challenges
9. Limited Research on Non-Infectious Diseases
10. Need for Booster Doses

16. Types of mRNA Vaccines
mRNA vaccines can be classified into three main types based on their mechanism of action and ability to amplify
mRNA within cells.
1. Conventional mRNA Vaccines (Non-Replicating mRNA Vaccines)
Mechanism:
•Contains a single-stranded mRNA that encodes the antigen (e.g., viral spike protein).
•After delivery into cells, the mRNA is translated by ribosomes into the antigenic protein.
•The antigen is then presented to the immune system, triggering an immune response.
•The mRNA does not replicate and is eventually degraded by cellular processes.

17. Features:
 Simple design with no need for additional replication mechanisms.
 Requires higher doses because the mRNA does not replicate itself.
 Easier to manufacture and widely used in commercial vaccines.
Pfizer-BioNTech and Moderna COVID-19 vaccines.

18. 2. Self-Amplifying mRNA (saRNA) Vaccines (Replicon mRNA Vaccines)
Mechanism:
•Contains two components:
• mRNA encoding the antigen (target protein).
• mRNA encoding viral replication machinery (usually from alphaviruses, like RNA-dependent RNA polymerase).
•Once inside the cell, the viral replication machinery amplifies the mRNA, producing multiple copies of the antigenic protein.
•This increases antigen production and enhances immune response with a lower initial mRNA dose.
Features:
✔ Higher efficiency -requires a lower dose than conventional mRNA vaccines.
✔ Longer-lasting antigen production, leading to a stronger immune response.
✔ Still in experimental stages – under development for flu, Zika, and cancer vaccines.
Example: Experimental saRNA COVID-19 vaccines by Imperial College London and Gritstone Bio.

19. 3. Trans-Amplifying mRNA (taRNA) Vaccines
Mechanism:
•Uses two separate mRNA molecules:
• Antigen-encoding mRNA (like in conventional vaccines).
• Helper mRNA that provides replication enzymes (but not the full viral machinery).
•Unlike self-amplifying mRNA, where the replication genes are on the same strand as the antigen, trans-amplifying
mRNA vaccines separate them, allowing flexibility in vaccine design.
Features:
 More controlled replication than self-amplifying mRNA, reducing unwanted side effects.
 Lower risk of immune responses against viral replication proteins, which is a concern in saRNA vaccines.
 Still in early research stages, with potential applications in infectious diseases and cancer therapy

21. The mRNA Vaccine Delivery Systems
Polymeric Nanoparticles
Peptides and Proteins Nanoparticles
Protamine Nanoparticles
Lipid Nanoparticles (LNPs)
Exosome-based systems

22. Polymeric Nanoparticles
Encapsulation: mRNA binds to cationic polymers for protection.
Cellular Uptake: Nanoparticles enter cells via endocytosis.
Endosomal Escape: pH-responsive polymers release mRNA into the cytoplasm.
Degradation: The polymeric carrier is broken down and cleared.
Targeting ligands can be
added to the polymeric
surface for cell-specific
delivery (e.g., targeting
dendritic cells or tumor
cells).
• Polyethyleneimine (PEI)
– Effective but can be
toxic.
• Poly(β-amino esters)
(PBAEs) – More
biodegradable and less
toxic than PEI.
• Chitosan Nanoparticle

23. Peptides and Proteins Nanoparticles
mRNA Encapsulation & Protection – Cationic
peptides (e.g., arginine-rich peptides) electrostatically
bind to negatively charged mRNA, forming stable
nano-complexes that protect mRNA from degradation.
Cellular Uptake (Endocytosis & Membrane
Fusion) – Peptide-mRNA complexes enter cells via
endocytosis, and pH-sensitive cell-penetrating
peptides (CPPs) like RALA peptides alter their
structure in acidic endosomes, facilitating membrane
fusion.
Endosomal Escape & mRNA Release – Peptides
disrupt endosomal membranes, allowing mRNA
release into the cytoplasm, preventing degradation and
enabling translation
repeated
arginine-
alanine-
leucine-
alanine

24. Protamine Nanoparticles
mRNA Encapsulation & Protection – Protamine, a cationic protein rich in arginine, electrostatically binds to
negatively charged mRNA, forming stable nano-complexes..
Cellular Uptake (Endocytosis) – The protamine-mRNA complex enters the cell via endocytosis.
Endosomal Escape & Cytoplasmic Release – Protamine disrupts endosomes, releasing mRNA into the cytoplasm

25. Lipid Nanoparticles (LNPs)
Encapsulation & Protection – LNPs shield mRNA from degradation.
Cellular Uptake – LNPs enter cells via endocytosis.
Endosomal Escape – Ionizable lipids disrupt endosomes, releasing mRNA into the cytoplasm.
Translation & Immune Activation – mRNA is translated into an antigen, triggering an immune response.

26. LNPs typically consist of:
✔ Ionizable lipids (40–50%) – Aid in mRNA
binding and endosomal escape.
✔ Cholesterol (38–45%) – Enhances membrane
stability.
✔ Helper phospholipids (10–12%) – Assist in
lipid bilayer fusion.
✔ PEGylated lipids (1–2%) – Improve circulation
time and prevent aggregation.

27. Microfluidics Process for LNP-mRNA Formulation
Controlled Mixing – mRNA (aqueous buffer) and lipids (ethanol) are precisely mixed in a microfluidic device.
Self-Assembly of LNPs – Lipids encapsulate mRNA, forming stable LNP-mRNA complexes with high efficiency.
Size Control & Homogeneous Formation – Microfluidic channels regulate LNP size (~50-100 nm) for consistent delivery.
Purification (Tangential Flow Filtration - TFF) – Removes excess ethanol and buffers, stabilizing the formulation.
Sterilisation & Final Product – The purified LNP-mRNA is filtered (0.2 μm) and prepared in liquid or lyophilized form

29. Encapsulation & Engineering
Cellular Uptake
Endosomal Escape
Translation & Immune Activation
Exosome-based systems
small extracellular vesicles (EVs) (30–150 nm) secreted by
cells that play a crucial role in cell-to-cell communication by
transporting proteins, lipids, RNA, and mRNA between cells.
They originate from endosomal compartments and are
released when multivesicular bodies (MVBs) fuse with the
plasma membrane

30. The mRNA Vaccine Applications
First mRNA therapy
was tested in 1992
(vasopressin-
encoding mRNA for
diabetes insipidus)
Infectious Disease Vaccines
Cancer Immunotherapy
Autoimmune & Rare Disease Therapies
Protein Replacement Therapy
Regenerative Medicine
Gene Editing & CRISPR Applications
Neurodegenerative Disease Therapies
Pandemic Preparedness & Biodefense

31. Infectious Disease Vaccines
Current mRNA Vaccines (Approved & In Use)
1.COVID-19 (SARS-CoV-2)
1. Pfizer-BioNTech (BNT162b2) & Moderna (mRNA-1273)
2. First-ever mRNA vaccines approved.
2.Influenza (Clinical trials – Phase 3)
1. Moderna’s mRNA-1010, targeting multiple flu strains.
Future mRNA Vaccines (Under Development)
3.Zika Virus – Preclinical & early-phase trials for high-risk areas.
4.Rabies – CureVac’s CV7202 shows promising immunogenicity.
5.Cytomegalovirus (CMV) – Moderna’s mRNA-1647 in clinical trials.
6.HIV – mRNA-1644 (Moderna & NIH) in Phase 1 trials.
7.Tuberculosis (TB) – Potential mRNA alternative to BCG vaccine.
8.Malaria – Targeting Plasmodium species, under development.
9.Dengue, Chikungunya, Ebola – Various early-stage trials.

32. Autoimmune & Rare Disease Therapies
1. Autoimmune Diseases
•Multiple Sclerosis (MS): mRNA therapy trains immune tolerance to myelin antigens.
•Type 1 Diabetes: mRNA helps protect and regenerate pancreatic beta cells.
•Inflammatory Bowel Disease (IBD): mRNA modulates immune response to reduce gut
inflammation.
2. Rare Genetic Disorders
•Cystic Fibrosis (CF): mRNA therapy restores CFTR protein function.
•Propionic Acidemia & Methylmalonic Acidemia: Corrects enzyme deficiencies.
•Lysosomal Storage Disorders (e.g., Gaucher, Pompe, Fabry disease): mRNA replaces
missing or defective enzymes.
•Glycogen Storage Diseases: mRNA restores glycogen metabolism enzymes.
(Ongoing Research & Clinical Trials)
reprogramming the
immune system to restore
immune tolerance and
prevent excessive immune
responses.
introducing specific
mRNA sequences that
encode tolerogenic
proteins or anti-
inflammatory factors
to regulate the
immune response.

33. Future Applications (Emerging Research & Potential Uses)
Autoimmune & Rare Disease Therapies
Personalized Autoimmune Therapies
•Tailored mRNA vaccines to modulate immune response for Rheumatoid Arthritis, Lupus, IBD and Psoriasis.
Tolerogenic mRNA Vaccines
•Teaches the immune system to recognize self-antigens, preventing autoimmune attacks.
•Potential in Multiple Sclerosis, Type 1 Diabetes, Myasthenia Gravis and Autoimmune Hepatitis.
Gene Editing & Protein Replacement Therapies
•Duchenne Muscular Dystrophy (DMD): mRNA increases dystrophin protein expression.
•Hemophilia: mRNA-based clotting factor replacement.

34. Cancer Immunotherapy
Personalized Cancer Vaccines
•mRNA-based neoantigen vaccines stimulate immune responses against patient-specific tumor mutations.
(Moderna & BioNTech’s mRNA cancer vaccines in clinical trials for melanoma, lung, and pancreatic cancer)
Off-the-Shelf Cancer Vaccines
•Targets common tumor-associated antigens (TAAs) like HER2 (breast cancer), MAGE-A3 (lung cancer), NY-ESO-1
(sarcoma & melanoma).

35. mRNA-Encoded Cytokine Therapy
• mRNA delivers cytokines (e.g., IL-2, IL-12, IFN-γ) to enhance anti-tumor immune response.
• Helps overcome immunosuppressive tumor microenvironments.
mRNA for Chimeric Antigen Receptor (CAR) T & NK Cell Therapy
• mRNA engineering of CAR-T and CAR-NK cells enhances their ability to target cancers.
• Avoids permanent genetic modifications, improving safety.
Cancer Immunotherapy

36. Protein Replacement Therapy
Monoclonal Antibody Production
mRNA instructs cells to produce therapeutic
monoclonal antibodies inside the body, reducing
the need for external antibody infusions.
Hormone Replacement Therapy
Insulin (Diabetes): mRNA-based insulin therapy
to restore glucose regulation.
Growth Hormone Deficiency: mRNA therapy for
controlled human growth hormone (hGH)
production.
Clotting Factor Therapy
(Hemophilia)
mRNA encodes Factor VIII or IX to
treat Hemophilia A and B, reducing
dependency on regular injections.
Erythropoietin (EPO) Therapy
(Anemia & Kidney Disease)
mRNA-based EPO production for
chronic kidney disease and
chemotherapy-induced anemia.

37. Protein Replacement Therapy
Lysosomal Storage Disease Therapy
mRNA-based replacement of missing
enzymes for Gaucher, Fabry, Pompe,
and Tay-Sachs diseases.
Cystic Fibrosis (CF) Therapy
mRNA restores CFTR protein function,
addressing the root cause of CF.
mRNA-Based Therapies for
Muscular Dystrophies
Duchenne Muscular Dystrophy (DMD):
mRNA therapy enhances dystrophin
production.
Enzyme Replacement for
Metabolic Disorders
mRNA encodes metabolic enzymes
for disorders like Phenylketonuria
(PKU) and Glycogen Storage
Diseases.
mRNA for Neurodegenerative
Disease Treatments
Alzheimer’s & Parkinson’s Disease:
mRNA restores neuroprotective
proteins to slow disease progression.
Self-Amplifying mRNA for Long-
Lasting Protein Therapy

38. Regenerative Medicine
• mRNA-based therapy to promote cardiac muscle repair after heart attacks
(myocardial infarction).
• Encodes growth factors like VEGF (vascular endothelial growth factor) to
stimulate new blood vessel formation.
Cardiac Regeneration (Heart
Disease Treatment)
• mRNA instructs cells to produce collagen and growth factors to accelerate
wound closure and tissue healing.
• Potential for burn treatment and diabetic wound repair.
Wound Healing & Skin
Regeneration
• mRNA delivers bone morphogenetic proteins (BMPs) to enhance bone and
cartilage repair.
• Studied for treating osteoporosis, arthritis, and non-healing fractures.
Cartilage & Bone
Regeneration (Osteoarthritis
& Fracture Healing)

39. • mRNA therapy to stimulate neural regeneration and promote functional recovery
in spinal cord injury patients.
Spinal Cord Injury Repair
• mRNA-driven therapies to repair liver damage caused by cirrhosis, liver failure,
or chronic diseases.
Liver Regeneration
• mRNA-encoded neurotrophic factors to regenerate neurons in
neurodegenerative diseases like Alzheimer’s, Parkinson’s, and stroke recovery.
Neuroregeneration (Brain & Nerve
Repair)
• mRNA instructs cells to regenerate insulin-producing beta cells, offering a
potential cure for Type 1 Diabetes.
Regeneration of Pancreatic Beta
Cells (Diabetes Treatment)
• mRNA encodes proteins to repair damaged lung tissue in chronic obstructive
pulmonary disease (COPD) and pulmonary fibrosis.
Lung Regeneration (Respiratory
Diseases)
• Converts adult cells into pluripotent stem cells (iPSCs) for personalized
regenerative medicine.
mRNA-Induced Stem Cell Therapy
Self-Amplifying mRNA (saRNA) for Long-
Term Regeneration

40. Neurodegenerative Disease Therapies
Alzheimer’s Disease (AD)
mRNA therapy encodes neuroprotective proteins (e.g., brain-derived neurotrophic
factor BDNF) to support neuron survival.
mRNA-based beta-amyloid and tau-targeting antibodies to slow disease
progression.
Parkinson’s Disease (PD)
mRNA therapy stimulates dopamine-producing neurons by delivering growth
factors like glial cell line-derived neurotrophic factor (GDNF).

41. Amyotrophic Lateral Sclerosis (ALS)
mRNA-based therapy targets superoxide dismutase 1 (SOD1) mutations, a major cause of ALS.
Encourages motor neuron survival and function.
Huntington’s Disease (HD)
mRNA therapy designed to reduce mutant huntingtin protein (mHTT) levels, preventing neuronal damage
mRNA for Stroke Recovery & Neural Regeneration
mRNA-driven therapies to stimulate angiogenesis and neurogenesis after ischemic stroke.

42. • Allows lower-dose vaccines with prolonged protection, ideal for mass
immunization during outbreaks.
Self-Amplifying mRNA (saRNA) for
Pandemic Response
• Developing mRNA-based countermeasures for anthrax, smallpox,
and hemorrhagic fever viruses (Ebola, Marburg, Lassa fever).
mRNA Vaccines for Bioterrorism
Threats
• WHO and global research groups using AI-driven mRNA design to
predict and prepare for unknown future pathogens.
mRNA Vaccines for High-Risk Viruses
("Disease X")
• Next-gen mRNA vaccines stored at room temperature, solving cold-
chain logistics issues.
Thermostable mRNA Vaccines for
Global Readiness
• On-demand mRNA therapies producing neutralizing antibodies
against new viral threats.
mRNA-Based Monoclonal Antibodies
for Rapid Defense
Combination mRNA Vaccines for
Multi-Pathogen Protection
Single-dose vaccines covering multiple
viruses (e.g., COVID-19 + Flu + RSV).
Pandemic Preparedness & Biodefense

43. mRNA vaccine technology in India
Gennova Biopharmaceuticals Ltd. (Pune)
•Developed GEMCOVAC®-19, India's first mRNA COVID-19 vaccine.
•Created GEMCOVAC®-OM, an Omicron-specific booster.
Serum Institute of India (SII) (Pune)
•Exploring mRNA-based cancer immunotherapy in collaboration with
Univercells (Belgium).
Centre for Cellular and Molecular Biology (CCMB) (Hyderabad)
•Developed India’s first indigenous mRNA vaccine technology with
CSIR

46. mRNA is no longer just about vaccines—it’s the future of medicine!