Provided herein are compositions comprising minigenes comprising splice modulator binding sequences, for regulatable gene expression, and systems and methods of use thereof.

Disclosed herein are cells expressing DUX4 including stem cells, differentiated cells thereof, primary T cells, and chimeric antigen receptor T cells, as well as related methods of their use and generation. In some embodiments, the cells disclosed herein do not express one or more MHCI and/or MHC II human leukocyte antigens. In some embodiments, such cells possess immune evasion properties.

The invention provides methods and compositions for perturbing gene expression in hematopoietic cell lineages in vivo.

The present invention relates generally to mutagenesis of target genes that takes advantage of the natural mutagenic capabilities of B cells, and enhances those capabilities by bringing the process of diversification under control. The invention provides a method for rapidly and inducibly generating point mutations and other types of diversification in expressed genes, such as antibody genes. This method can be coupled with selection to identify B cell clones that produce, for example, antibodies of high affinity or specificity. The diversification process can be modulated, accelerated, halted, switched between methods of mutagenesis and the like. The modulation of diversification in accordance with the invention is both inducible and reversible. The invention provides a means of rapid and feasible development of a repertoire of variant immunoglobulins and other polypeptides.

The invention provides a platform and methods of using the platform for the regulation of the expression of a target gene using exposure to an aptamer ligand (for example, a small molecule). The platform features a polynucleotide gene regulation cassette that is placed in the target gene and includes a synthetic riboswitch positioned in the context of a 5′ intron-alternative exon-3′ intron. The riboswitch comprises an effector region and a sensor region (e.g., an aptamer that binds a small molecule ligand) such that the alternative exon is spliced into the target gene mRNA when the ligand is not present thereby preventing expression of the target gene. When the ligand is present, the alternative exon is not spliced into the target gene mRNA thereby providing expression of the target gene.

The present disclosure provides an in vitro mammalian cell that is genetically modified to provide for enhanced production of adeno-associated virus (AAV) virions. The mammalian cells can be used to produce AAV virions, e.g., recombinant AAV virions that include a heterologous nucleic acid encoding a gene product; the present disclosure thus provides methods for producing an AAV virion, which may be a recombinant AAV virion.

The presently disclosed subject matter relates to targeted integration (TI) host cells suitable for the expression of recombinant proteins wherein those TI host cells have been subjected to supertransfection resulting in the random integration (RI) of exogenous nucleic acids encodes into their genome, as well as methods of producing and using said supertransfected TI host cells.

The present invention relates to viral particles which exhibit self-regulatory or regulatable features.

The present inventions generally relate to site-specific delivery of nucleic acid modifiers and includes novel DNA-binding proteins and effectors that can be rapidly programmed to make site-specific DNA modifications. The present inventions also provide synthetic all-in-one genome editor (SAGE) systems comprising designer DNA sequence readers and a set of small molecules that induce double-strand breaks, enhance cellular permeability, inhibit NHEJ and activate HDR, as well as methods of using and delivering such systems.

The present invention provides genetically modified microorganisms with reduced transmissibility, wherein the microorganism has been genetically modified to express a single stranded binding protein (SSB) regulated by an arabinose responsive promoter, wherein the microorganism has a reduced growth capacity in the absence of arabinose.