Mercers team recognized benefits of cDNA-based long-read nanopore sequencing for determining mRNA vaccine integrity.A well-earned spotlight shines on mRNA vaccines, with recent Nobel prize accolades awarded to Katalin Karikó and Drew Weissman for their fundamental research that made it possible for effective and fast vaccine advancement during the covid pandemic. Researchers outside the vaccine production field regularly utilize sequencing to examine elements of DNA and RNA biology, consisting of gene expression, RNA polyadenylation or poly(A) tails, and post-transcriptional chemical modifications that impact RNA stability and immune activation.2 In Mercers latest work published in Nature Communications, his group established a structured approach called VAX-seq to examine mRNA vaccines using long-read nanopore sequencing.3 Their technique measures key vaccine quality associates at various production actions, consisting of sequence contamination.the, integrity, and identity scientists examined and compared several sequencing methods, consisting of brief- and long-read sequencing of cDNA that they created from the mRNA vaccine and direct RNA sequencing with long-read nanopore innovation. Mercers team effectively analyzed vaccine sequences with each strategy and determined cDNA-based long-read sequencing as the best approach for measuring vaccine quality.”See Also “Newly Developed mRNA Vaccine Protects Against Lyme Disease”Although Mercers group found cDNA sequencing more trustworthy than direct RNA sequencing, they anticipate that long-read RNA sequencing will continue to be a useful research tool for mRNA vaccine development. Gunter HM, et al. mRNA vaccine quality analysis utilizing RNA sequencing.
Mercers team recognized advantages of cDNA-based long-read nanopore sequencing for determining mRNA vaccine integrity.A well-earned spotlight shines on mRNA vaccines, with current Nobel reward awards awarded to Katalin Karikó and Drew Weissman for their fundamental research study that made it possible for fast and efficient vaccine development during the covid pandemic. As mRNA vaccine research study gains ground in applications beyond covid immunization, tools that allow scientists to improve their production approaches are increasingly crucial. Until recently, vaccine producers mainly depended on techniques such as gel electrophoresis and liquid chromatography, which researchers initially embraced for protein-based vaccines and pharmaceuticals that held center stage prior to the pandemic.1 Although dependable and robust, these quality assurance methods are lengthy and pricey for analyzing mRNA vaccine products. “During the pandemic, certainly since of the sped up speed, a great deal of those processes were just inherited from standard maker workflows,” described Timothy Mercer, an RNA biologist at the University of Queensland who leads an mRNA manufacturing center developed on artificial biology, genomics, and transcriptomics research. “The actual item itself got moved a lot faster than the environment around it, which is now in the process of capturing up.” See Also “Nobel Prize for mRNA Vaccines”The actual item itself got moved a lot faster than the ecosystem around it, which is now in the procedure of catching up.-Timothy Mercer, University of QueenslandFor Mercer, bringing sequencing approaches into that community was an instinctive solution for improving quality assurance analyses. Scientists outside the vaccine manufacturing field regularly use sequencing to analyze elements of DNA and RNA biology, consisting of gene expression, RNA polyadenylation or poly(A) tails, and post-transcriptional chemical adjustments that affect RNA stability and immune activation.2 In Mercers most current work published in Nature Communications, his team established a structured method called VAX-seq to analyze mRNA vaccines using long-read nanopore sequencing.3 Their approach measures crucial vaccine quality associates at different manufacturing actions, including sequence contamination.the, stability, and identity scientists examined and compared several sequencing methods, consisting of short- and long-read sequencing of cDNA that they produced from the mRNA vaccine and direct RNA sequencing with long-read nanopore innovation. Mercers team effectively examined vaccine sequences with each method and recognized cDNA-based long-read sequencing as the very best approach for determining vaccine quality. Unlike short-read approaches, long-read nanopore sequencing was efficient for full-length and fragmented mRNAs, which supplied insight into mRNA degradation. “Because youre sequencing the entire particle, youre not simply determining what the series of that particle is, but youre also saying its a full-length practical molecule,” Mercer said. “Thats an additional quality metric, which is truly key to the real pharmaceutical activity of that specific mRNA sample.” This advantage lines up with long-read sequencings recognized strengths. “I believe that the techniques theyve developed are extremely existing and of interest.” stated Cynthia Burrows, a chemist from the University of Utah who is a nucleic acids professional and who was not associated with the research study. “As a researcher, what you would like to know is, are the majority of your strands complete length or has there been some aborted synthesis? Does it all have the poly(A) tail at the end? These are the sorts of things that the paper resolved.”See Also “Newly Developed mRNA Vaccine Protects Against Lyme Disease”Although Mercers team found cDNA sequencing more reputable than direct RNA sequencing, they expect that long-read RNA sequencing will continue to be a useful research tool for mRNA vaccine advancement. Burrows and other scientists have actually currently used direct RNA sequencing to examine the residential or commercial properties of nucleoside modifications,4 such as the pseudouridine adjustment in mRNA vaccines that amassed Katalin Karikós and Drew Weissmans Nobel acknowledgment. “That was so far ahead of its time, and the fights that were fought there to keep moving on and keep the science going so that all of a sudden, when we truly required mRNA vaccines, they were there for us. That is an incredible story,” Burrows said. Sequencing mRNA vaccines is another important chapter because story, moving the science forward by linking technical developments with translational breakthroughs. Helen Gunter, the postdoctoral fellow in Mercers laboratory who led this work, is already broadening VAX-seq previous quality assurance applications by teaming up with other research laboratories throughout Australia. “Shes starting to get an actually mutual understanding throughout all these different model systems, all these various cells, about the diversity of methods which the input, the RNA, is in fact taken up into the cell, processed, and expressed,” said Mercer. “I believe originating from that biological viewpoint, the sequencing is an actually beneficial tool.”ReferencesRosa SS, et al. mRNA vaccines making: Challenges and traffic jams. Vaccine. 2021; 39( 16 ):2190 -200. Baronti L, et al. A guide to large-scale RNA sample preparation. Anal Bioanal Chem. 2018; 410( 14 ):3239 -52. Gunter HM, et al. mRNA vaccine quality analysis using RNA sequencing. Nat Commun. 2023; 14( 1 ):5663. Fleming AM, Burrows CJ. Nanopore sequencing for N1-methylpseudouridine in RNA reveals sequence-dependent discrimination of the customized nucleotide triphosphate throughout transcription. Nucleic Acids Res. 2023; 51( 4 ):1914 -26.
