Abstract: A manufacturing process of RNA having a length of about 20-200 bases with improved performance, by using in vitro transcription in combination with other methodologies that may increase yield and quality.

Abstract: The invention provides, inter alia, improved replicons and vectors encoding them, where the replicons provide sustained expression of an encoded protein. These replicons comprise flavivirus replicases and heterologous protein coding sequences, wherein the heterologous protein coding sequences are flanked by separation sequences for improved efficacy. These nucleic acids provided by the invention, including self-replicating RNAs provided by the invention, are useful in methods of protein expression, such as for vaccines (e.g., for methods of immunization), as well as expression of therapeutic proteins, such as antibodies (e.g., for methods of treatment).

Abstract: Described herein are methods of analyzing and producing RNA compositions, e.g., mRNA compositions, that include determining the amount of a polyA chain length and/or 5 the relative distribution of polyA chain lengths in a sample from the RNA composition using mass spectrometry, e.g., LC-MS or MALDI-MS.

Abstract: The invention provides, inter alia, improved replicons and vectors encoding them, where the replicons provide sustained expression of an encoded protein. These replicons comprise flavivirus replicases and heterologous protein coding sequences, wherein the heterologous protein coding sequences are flanked by separation sequences for improved efficacy. These nucleic acids provided by the invention, including self-replicating RNAs provided by the invention, are useful in methods of protein expression, such as for vaccines (e.g., for methods of immunization), as well as expression of therapeutic proteins, such as antibodies (e.g., for methods of treatment).

Abstract: Described herein are recombinant vaccinia viruses that provide the efficacy of the current smallpox vaccine, but with a built-in safety mechanism, giving the physician or vaccine recipient (vaccinee) control over the vaccine virus replication. Specifically, genetic elements of the tetracycline (tet) operon and modified tet repressor genes are used to control the expression of vaccinia virus genes that are essential for viral replication, thereby allowing replication of the virus to be accurately regulated through the addition or removal of antibiotics (tetracyclines). The recombinant vaccinia viruses can be used as safer next-generation smallpox vaccines, as expression vectors for exogenous genes such as those encoding therapeutic or toxic proteins, as oncolytic viruses, and for tumor imaging and vector tracking in vivo.