Thom Aldrin

CureVac patent WO2016180430A1, as cited in US20210261897, primarily concerns a method for producing RNA (particularly mRNA) by in vitro transcription, using DNA (mainly plasmid DNA) as a template. WO2016180430A1 patents.google.com/patent/WO20161… The following information was extracted by Grok. Disadvantages/Risks of DNA in Therapeutic Applications (Compared to RNA) The patent explicitly lists several advantages of RNA over DNA, thereby highlighting the following drawbacks of DNA: • Risk of genomic integration and mutation:DNA can integrate into the host cell genome to a certain degree and may insert into an intact gene, causing mutations that lead to partial or complete loss of genetic information or misinformation. RNA does not integrate into the genome. • Requirement for strong (viral) promoters and associated side effects: DNA expression requires strong promoters (e.g., viral CMV promoter). If such promoters integrate into the genome, they can cause undesirable changes in the regulation of host gene expression. RNA transcription does not require viral sequences or promoters. • Non-transient (permanent) expression: Once integrated, DNA leads to persistent expression that cannot be discontinued after the treatment period. RNA enables transient gene expression because it degrades within a limited time. • Induction of undesirable immune responses: DNA can induce pathogenic anti-DNA antibodies, causing unwanted immune reactions. RNA does not lead to the induction of pathogenic anti-RNA antibodies. • Limited applicability and slower preparation:RNA is widely applicable and can be prepared quickly for any desired protein, even on an individual patient basis (personalized medicine). DNA has a narrower range of applications and is less suitable for rapid preparation. Disadvantages in Manufacturing and Quality Control (as Template DNA) DNA (especially plasmid DNA amplified in E. coli) serves as the necessary template for in vitro transcription but introduces several challenges: • High contamination risk: Residual template DNA (or bacterial genomic DNA) can contaminate the final RNA product. Complete removal is essential and typically requires additional steps such as DNase digestion, which complicates the process. • Various impurities from bacterial production: • Endotoxins (lipopolysaccharides from gram-negative bacteria): Must be strictly minimized due to immunotoxicity risks. • Residual bacterial (e.g., E. coli) genomic DNA. • Proteins, RNA contaminants, and RNases (ribonucleases) that can inhibit transcription or degrade the RNA product. • Bioburden (microbial load). • Complex and burdensome quality control: This increases process complexity and the risk of batch-to-batch variability. Template DNA requires extensive testing for concentration, integrity, identity, and purity, including: • Photometric measurement (e.g., OD260/280). • PCR or qPCR for identity and residual bacterial DNA. • Restriction enzyme digestion + agarose gel electrophoresis (for linearization confirmation and identity). • RNase treatment to assess RNA contamination. • LAL test for endotoxins. • Protein quantification (e.g., BCA assay). • Sequencing. • Linearization challenges: Circular plasmid DNA must be linearized with restriction enzymes for proper transcription termination. Incomplete linearization can lead to undefined RNA transcripts, reduced yield, or quality issues. • Difficulties in large-scale cGMP/GMP production: Amplification in bacteria involves fermentation, purification, and contamination control. No fully established cGMP-compliant large-scale process for high-purity DNA templates existed at the time, making consistent production challenging. Harsh conditions (organic solvents, high temperatures) may be needed in some purification steps. • Mutation risk in certain amplification methods: PCR-based synthesis of template DNA can introduce more mutations compared to plasmid cloning and amplification in bacteria. @Jikkyleaks @Kevin_McKernan 📷

La maladie de Lyme : des documents déclassifiés américains relient l’épidémie à des programmes d’armes biologiques En France, c 'est l'omerta pgibertie.com/2026/04/14/la-… via @GibertiePatrice

La justice française refuse d’exécuter une commission rogatoire internationale de la Suisse, laquelle demande la saisie des biens des Moretti, les propriétaires du bar de la station de ski Crans-Montana ➡️ 20min.fr/j05

Parents, here’s something you may not know: In 1954, a massive polio vaccine trial followed about 200,000 vaccinated children and 200,000 who received a placebo—and found no significant difference in nonparalytic polio rates between the two groups. And to date, it hasn’t been proven that the IPV (polio) vaccine is safer than polio itself. Get the referenced facts in one place: picdata.org/polio-dis/ #parenting