The invention provides in vivo methods for identifying cancer-associated immunotherapy targets.

The present disclosure provides nucleobase editors that include a cytidine deaminase domain, a codon-optimized nuclease-defective Cas9 domain, and at least one nuclear-localization sequence. The nucleobase editors disclosed herein improve the efficiency by which single-nucleotide variants can be created compared to conventional BE3 nucleobase editors.

Particular aspects provide for use of the β-herpesvirus Cytomegalovirus (CMV: e.g., RhCMV and HCMV) as a uniquely evolved “vector” for safely initiating and indefinitely maintaining high level cellular and humoral immune responses (against, e.g., HIV, SIV, TB, etc.). Particular aspects provide a method for treatment or prevention of, e.g., HIV, SIV or TB, comprising infection of a subject in need thereof with at least one recombinant CMV-based vector (e.g., HCMV or RhCMV) comprising an expressible HIV/SIV/TB antigen or a variant or fusion protein thereof. In particular embodiments of the method, infection is of an immunocompetent, HCMV or RhCMV seropositive subject. Additional aspects provide for RhCMV- and HCMV-based vaccine vectors, and versions thereof with suicide or safety means. Further aspects provide pharmaceutical compositions comprising the inventive CMV-based vaccine vectors.

The present specification relates to a transgenic animal and a transgenic embryo producing components of an engineered nuclease.

According to the disclosure of the present specification, the transgenic animal (or embryo) producing components of an engineered nuclease is a transgenic animal (or embryo) which includes a first cell having a genome including a first toolbox; and a second cell having a genome including a second toolbox, wherein the first toolbox and the second toolbox include at least one of a polynucleotide encoding an RNA-guided endonuclease and a polynucleotide encoding a guide nucleic acid that is able to specifically bind to a target site, respectively, wherein the first toolbox is present in a first locus of the genome of the first cell; the second toolbox is present in a second locus of the genome of the second cell; and the first locus is different from the second locus.

The invention provides inducible promoter systems and their components incorporating components of a tetracycline operon. By coordinating expression of different transcriptional units in these systems as a result of selection of promoters and/or linking the units into the same DNA molecule, these systems can achieve higher levels of expression of coding segments of interest, increased differential levels of expression between on- and off-states, and/or greater responsiveness to inducing agents than conventional systems.

A trophoblast stem cell-like cell having potential to differentiate into a placenta-constituting cell which is induced from a trophoblast cell derived from a placenta of or after the second trimester of pregnancy and which contains a SALL4 gene functionally linked to a first exogenous inducible promoter. Also, a method for producing a trophoblast stem cell-like cell having potential to differentiate into a placenta-constituting cell.

Described herein are cell lines for recombinant adeno-associated virus (AAV) production, cell lines for AAV-implemented protein production, and cell lines for use in titering AAV; methods of making each of those cell lines; and methods of using each of those cell lines. In some aspects, the cell lines disclosed herein may be used in a manufacturing process that is seed virus-free, helper virus-free, and transfection-free that uses synthetic elements to control viral genes in a stable cell line.

The invention relates to retroviral producer cell comprising nucleic acid sequences encoding: gag and pol proteins; envelope protein or a functional substitute thereof; amplifiable selection marker; and the RNA genome of the retroviral vector particle, wherein said nucleic acid sequences are all integrated at a single locus within the retroviral producer cell genome. The invention also relates to nucleic acid vectors comprising a non-mammalian origin of replication and the ability to hold at least 25 kilobases (kb) of DNA, characterized in that said nucleic acid vector comprises retroviral nucleic acid sequences encoding: gag and pol proteins, and an env protein or a functional substitute thereof. The nucleic acid vector additionally comprises nucleic acid sequences encoding an amplifiable selection marker. The invention also relates to uses and methods using said nucleic acid vector in order to produce stable retroviral packaging and producer cell lines.

Aspects of this disclosure provide nucleic acid constructs A nucleic acid construct, comprising a first expression cassette comprising a nucleic acid encoding a tetracycline-responsive transactivator under the control of a cell type-specific promoter; and a second expression cassette comprising a nucleic acid encoding a transgene, for example, a chimeric antigen receptor (CAR), under the control of a tetracycline-responsive promoter.

The invention provides a splice control module for sex-specific splicing and expression of a gene of interest. In certain embodiments, a dsx-based splice control module is used to express a lethal gene in an insect that is spliced in a sex-specific manner to impart lethality to female insects but not male insects.