ENAapplication/xmlftp://primaryOK200GenomicsINTERNATIONAL INSTITUTE OF MOLECULAR AND CELL BIOLOGY IN WARSAWhttps://www.ebi.ac.uk/ena/browser/view/PRJEB53190Despite the widespread use of mRNA vaccines against COVID-19 little is known about how therapeutic RNAs are metabolized in vivo. Here, we have implemented nanopore sequencing to analyze individual therapeutic mRNA molecules, providing information about poly(A) tails. We show that Moderna mRNA-1273 vaccine is terminated by ~100 nucleotide long poly(A) tail followed by a mΨCmΨAG sequence. In cell lines, mRNA-1273 is rapidly degraded in a process initiated by ΨCmΨAG removal, followed by CCR4-NOT-mediated deadenylation. However, mRNA-1273 poly(A) tails can be extended by up to 200 nucleotides in more medically relevant preclinical models, particularly in macrophages. Re-adenylation, which increases the stability of mRNA-1273, is mediated by TENT5A poly(A) polymerase and is dependent on TENT5A attachment to the endoplasmic reticulum (ER), where these mRNAs are translated. Intriguingly, BioNTech/Pfizer BNT162b2 does not undergo as potent re-adenylation as mRNA-1273 despite the similarity and ER targeting. Notably, in vivo, TENT5A is expressed in immune cells that take up mRNA-1273 after intramuscular administration. Furthermore, TENT5A deficiency reduces antigen and specific immunoglobulin production post-immunization in mice. Overall, our findings provide an unexpected principle for enhancing the efficacy of therapeutic mRNAs,opening up new opportunities for improvement.xref:PubMed:40102414xref:PubMed:40240603xref:EuropePMC:PMC12095053ENACOVID19, Policies, Vaccines, COVID-19, Utility, 2019 novel coronavirus, SARS CoV 2 Virus, COVID19 Virus, Easterlin, mRNA Vaccine, SARS, Novel Coronavirus, Virus, Antigen, β-coronavirus, Immune., taRNA Vaccine, Naked RNA Vaccines, Messenger RNA Vaccines, Microeconomic, 2019-nCoV, saRNA Vaccine, 2019 Novel Coronaviruses, Immune Processes, SARS Coronavirus 2, Theory, Immune Responses, β-CoVs, Utility Theories, Viruses, Household Consumption, Wuhan Seafood Market Pneumonia Virus, Economic Policies, severe acute respiratory syndrome coronavirus 2, Microeconomic Factors, Living Costs, Economic Conditions, beta-CoVs, RNA Vaccine, COVID-19 Virus, Immune Response, Immune, Capital, Household Consumptions, Economic, Theories, taRNA, Immune Process, Wuhan Coronavirus, Policy, Economic Policy, RNA, SARS-CoV-2 Viruses, 2019 Novel Coronavirus, β-CoV, RNA Vaccines, Home Economics, Indices, beta-CoV, Index, Process, COVID-19 Viruses, Conditions, Hypothesis, SARS-CoV-2, Coronavirus 2, Factor, Production, Living Cost, Macroeconomic Factors, Consumption, Economics, COVID19 Viruses, Economic Condition, Severe Acute Respiratory Syndrome Coronavirus 2, Household, Consumer Price Index, Consumer Price, SARS-CoV-2 Virus, Utility Theory, Coronavirus, Macroeconomic, Self-Amplifying RNA, 2019 Novel, betacoronavirus, Factors, mRNA, Coronavirus Disease 2019 Virus, Consumer Price Indices, Wuhan, Home, SARS-coronavirus 2, Self-Amplifying RNA Vaccine, Condition, Trans Amplifying RNA Vaccine, Easterlin Hypothesis, Remittance, Vaccine, Self Amplifying RNA Vaccine, Cost of Living, Response, 2019, COVID 19 Virus, saRNA, Self-Amplifying, RemittancesCOVID19, Policies, Vaccines, COVID-19, Utility, 2019 novel coronavirus, SARS CoV 2 Virus, COVID19 Virus, Easterlin, mRNA Vaccine, SARS, Novel Coronavirus, Virus, Antigen, β-coronavirus, Immune., taRNA Vaccine, Naked RNA Vaccines, Messenger RNA Vaccines, Microeconomic, 2019-nCoV, saRNA Vaccine, 2019 Novel Coronaviruses, Immune Processes, SARS Coronavirus 2, Theory, Immune Responses, β-CoVs, Utility Theories, Viruses, Household Consumption, Wuhan Seafood Market Pneumonia Virus, Economic Policies, severe acute respiratory syndrome coronavirus 2, Microeconomic Factors, Living Costs, Economic Conditions, beta-CoVs, RNA Vaccine, COVID-19 Virus, Immune Response, Immune, Capital, Household Consumptions, Economic, Theories, taRNA, Immune Process, Wuhan Coronavirus, Policy, Economic Policy, RNA, SARS-CoV-2 Viruses, 2019 Novel Coronavirus, β-CoV, RNA Vaccines, Home Economics, Indices, beta-CoV, Index, Process, COVID-19 Viruses, Conditions, Hypothesis, SARS-CoV-2, Coronavirus 2, Factor, Production, Living Cost, Macroeconomic Factors, Consumption, Economics, COVID19 Viruses, Economic Condition, Severe Acute Respiratory Syndrome Coronavirus 2, Household, Consumer Price Index, Consumer Price, SARS-CoV-2 Virus, Utility Theory, Coronavirus, Macroeconomic, Self-Amplifying RNA, 2019 Novel, betacoronavirus, Factors, mRNA, Coronavirus Disease 2019 Virus, Consumer Price Indices, Wuhan, Home, SARS-coronavirus 2, Self-Amplifying RNA Vaccine, Condition, Trans Amplifying RNA Vaccine, Easterlin Hypothesis, Remittance, Vaccine, Self Amplifying RNA Vaccine, Cost of Living, Response, 2019, COVID 19 Virus, saRNA, Self-Amplifying, RemittancesfalseSARS-CoV-2 mRNA vaccine is re-adenylated in vivo, enhancing antigen production and immune responseSARS-CoV-2 mRNA vaccine is re-adenylated in vivo, enhancing antigen production and immune response2025-04-282024-09-30PRJEB531904024060340102414