Methods are provided for transfecting cells with large cargo using a poly(beta-amino ester) (PBAE) molecule, and achieving high efficiency and viability. A method is provided of transfecting cells with a cargo, by forming a complex of the cargo with a (PBAE) molecule, mixing the complex with a first buffer and contacting the complex with the cells, wherein the cargo has a dimension of at least 0.1 μm. The PBAE molecule may be formed by reacting an amine with a di(acrylate ester). In some aspects, the PBAE molecule is poly(1,4-butanediol diacrylate-co-4-amino-1-butanol). In some aspects, the PBAE molecule is capped with 1-(3-aminopropyl)-4-methylpiperazine.

Recombinant adenovirus genomes that include a synthetic transcriptional circuit are described. Synthetic adenoviruses positively regulated using two-step transcriptional amplification (TSTA) are further described. Selection of the heterologous promoter is based on the desired replication characteristics of the synthetic virus. For example, the heterologous promoter can be a constitutive promoter, a tumor-specific promoter or a tissue-specific promoter.

Methods for rapidly confirming production of infectious viral vectors, for use in clinical grade personalized neo-antigen vaccines for subjects in need thereof, are provided.

The present disclosure provides a Farmington virus formulated to induce an immune response in a mammal against a tumour associated antigen. The Farmington virus may express an antigenic protein that includes an epitope from the tumour associated antigen. The Farmington virus may be formulated in a composition where the virus is separate from an antigenic protein that includes an epitope from the tumour associated antigen. The present disclosure also provides a prime:boost therapy for use in inducing an immune response in a mammal. The boost includes a Farmington virus, or a composition that includes a Farmington virus.

A vaccine for preventing β-CoV infection includes at least one viral vector containing a β-CoV DNA sequence which codes the S protein for the β-CoV. The β-CoV RNA sequence can be a SARS-2 β-CoV DNA sequence. The vaccine may further includes a packaging plasmid based on an adenovirus. The viral vector and packaging plasmid can be contained in a packaging cell and encapsidated in a capsid. A method of vaccinating a mammal subject against infection from at least one group of β-CoV includes separating a broad group of β-CoV into homology groups based on similarities in the β-CoV RNA sequences which code for their S proteins, identifying at least one consensus sequence for each homology group which has a sequence identity of greater than 60% to all other members of the homology group, and preparing a viral vector including at least a portion of the consensus sequence from at least one homology group.

The present disclosure provides chimeric adenoviral vectors comprising a nucleic acid encoding a coronavirus disease 2019 (COVID-19) protein and an adjuvant and methods for using the vectors to elicit an immune response to the SARS-CoV-2 protein in order to treat COVID-19.

The present invention relates to the preparation of mesenchymal stem cells, which are for the proliferation of viral vectors and viruses, and enable virus production and release timing control such that tumor targeting can be improved and viruses can be mass-produced. In addition, the tumorigenesis of mesenchymal stem cells of the present invention can be fundamentally blocked, safety is secured by enabling virus production and release to be controlled at desired time points, and antitumor effects can be maximized.

Methods for inducing an immune response against Human Immunodeficiency Virus (HIV) in HIV-infected subjects undergoing antiretroviral therapy (ART) are described. The methods involve initial administration of an adenovirus vector vaccine and subsequent administration of a poxvirus vector vaccine, followed by administration of anti-HIV broadly neutralizing antibodies (bNAb).

The present invention provides a recombinant adipose-derived stem cell and a recombinant method thereof, and belongs to the technical field of genetic engineering, where an adenovirus carrying an hFIX gene is transfected into an adipose-derived stem cell to obtain the recombinant adipose-derived stem cell. In the present invention, an adenovirus carrying an hFIX gene is transfected into an adipose-derived stem cell, and the recombinant adipose-derived stem cell obtained after the transfection can express an hFIX protein.

Systems and methods are presented that allow for determination and prediction of payload toxicity in therapeutic viruses. Disclosed herein are methods of determining payload toxicity of an expressed polypeptide in a cell, comprising: generating or procuring a plurality of expression vectors, each containing a different recombinant nucleic acid sequence that encodes a corresponding recombinant polypeptide; expressing the recombinant nucleic acid sequence in a plurality of host cells while culturing the host cells; sequencing the plurality of expression vectors after culturing the host cells; and correlating at least portions of the recombinant nucleic acid sequence with a toxicity measure.