Abstract: The invention features methods and apparatus for concentrating a nucleic acid. Methods of the invention include providing an initial suspension of a nucleic acid and an initial liquid, contacting the initial suspension with a housing having a filter that does not pass the nucleic acid, pressurizing the housing to produce a filtrate and a nucleic acid retentate from the initial solution, and detecting the volume of the nucleic acid retentate. Apparatus of the invention include a chamber configured to hold a filter housing containing a nucleic acid suspension, a pressure source to filter the suspension, and a detector to depressurize the housing upon detecting the volume reaching a predetermined threshold. The methods and apparatus described herein are useful in filtering, concentrating, and reconstituting nucleic acids, such as mRNA, in processes such as complete buffer replacement.

Abstract: Provided herein are methods of detecting mRNA purity in a mixture and related constructs. Also provided are compositions for separating and detecting full length mRNA from a mixture.

Abstract: RNA polymerase variants enable high efficiency transcription of RNA. In Yield some embodiments, the RNA polymerase variants enable RNA transcription with high capping efficiency and/or low levels of double-stranded RNA contamination.

Abstract: Provided herein are methods for detecting and/or measuring mRNA associated with ribosomes in a cell. The method can be used to assess potency and other characteristics of an miRNA drug product. Also provided are related products, including cells and reagents.

Abstract: Provided are formulations, compositions and methods for delivering biological moieties such as modified nucleic acids into cells to modulate protein expression. Such compositions and methods include the delivery of biological moieties, and are useful for production of proteins.

Abstract: The disclosure relates to HCMV ribonucleic acid (RNA) vaccines, as well as methods of using the vaccines and compositions comprising the vaccines.

Abstract: Provided herein are methods and compositions of a surrogate mRNA drug product. The surrogate mRNA drug product is made up of a physical property surrogate, a visual property surrogate, and a chemical property surrogate. The surrogate mRNA drug product has at least one quantitative or qualitative property such as viscosity, turbidity, density, and surface tension that is comparable to and matches the same property of a mRNA drug product comprised of mRNA in a lipid nanoparticle (LNP). The surrogate mRNA drug product, however, does not comprise mRNA or LNP.

Abstract: The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.

Abstract: The disclosure features novel methods of producing nucleic acid lipid nanoparticle (LNP) compositions employing a modifying agent after formation of a precursor nucleic acid lipid nanoparticle, the produced compositions thereof, and methods involving the nucleic acid lipid nanoparticles useful in the delivery of therapeutics and/or prophylactics, such as a nucleic acid, to mammalian cells or organs to, for example, to regulate polypeptide, protein, or gene expression.

Abstract: The present disclosure provides, in some aspects, variant RNA polymerases, the use of which increases transcription efficiency while reducing the number of double-stranded RNA contaminates and run-on transcripts produced during an in vitro transcription reaction.

Abstract: The invention relates to compositions and methods for the preparation, manufacture, and therapeutic use of ribonucleic acid vaccines comprising polynucleotide molecules encoding one or more antigens.

Abstract: The disclosure provides RNA vaccines for seasonal influenza virus as well as methods of using the vaccines.

Abstract: Aspects of the disclosure relate to nucleic acid vaccines. The vaccines include at least one RNA polynucleotides having an open reading frame encoding at least one varicella zoster virus (VZV) antigen. Methods for preparing and using such vaccines are also described.

Abstract: The present disclosure provides RNA polymerase variants for high efficiency transcription.

Abstract: The disclosure features novel lipids and compositions involving the same. Lipid nanoparticles (e.g., empty LNPs or loaded LNPs) include a novel lipid as well as additional lipids such as phospholipids, structural lipids, and PEG lipids. Lipid nanoparticles (e.g., empty LNPs or loaded LNPs) further including therapeutic and/or prophylactics such as RNA are useful in the delivery of therapeutic and/or prophylactics to mammalian cells or organs to, for example, regulate polypeptide, protein, or gene expression.

Abstract: The present disclosure in part provides compounds (i.e., PEG lipids) which are useful in pharmaceutical compositions, cosmetic compositions, and drug delivery systems, e.g. for use in lipid nanoparticle (LNP) formulations. The present disclosure also provides LNP formulations comprising PEG lipids described herein, and methods of using the same. For example, the LNPs provided herein are useful for the delivery of an agent (e.g., therapeutic agent) to a subject. The PEG lipids and LNPs provided herein, in certain embodiments, exhibit increased PEG shedding compared to existing PEG lipids and LNP formulations.

Abstract: The invention relates to concatemeric peptide epitope RNAs, as well as methods and compositions thereof. mRNA vaccines are also provided according to the invention, including cancer vaccines.

Abstract: The disclosure relates to cancer ribonucleic acid (RNA) vaccines, as well as methods of using the vaccines and compositions comprising the vaccines. In particular, the disclosure relates to concatemeric mRNA cancer vaccines encoding several cancer epitopes on a single mRNA construct, i.e. poly-epitope mRNA constructs or poly-neo-epitope constructs. The disclosure further relates to p53 and KRAS mutations, as well as incorporation of immune enhancers such as STING, e.g. mRNA constructs further encoding an immune stimulator or adjuvant. The disclosure further relates to inclusion of universal T cell epitopes, such as tetanus or diphtheria toxins to elicit an enhanced immune response.

Abstract: Aspects of the disclosure relate to methods of purifying mRNA by precipitating the mRNA, washing the precipitate to remove impurities and salts, and resuspending the washed mRNA to produce an mRNA composition. The disclosure describes reagents and methods useful for precipitation, washing, and resuspension of mRNA, and compositions produced by the methods described herein.

Abstract: The disclosure relates to ribonucleic acid (RNA) vaccines and combination vaccines, as well as methods of using the vaccines and compositions comprising the vaccines. Specific embodiments relate to RNA betacoronavirus vaccines formulated in a lipid nanoparticle.