The invention provides compositions and methods for delivering first and second polynucleotides each encoding a fragment of an A-to-G Base Editor fusion protein comprising one or more deaminases (e.g., adenosine deaminases) and nCas9, wherein the first polynucleotide encodes an N-terminal fragment of nCas9 fused to an intein-N of a split intein pair and the second polynucleotide encodes a C-terminal fragment of nCas9 fused to an intein-C of a split intein pair, and methods for delivering these fragments together with an sgRNA to a cell (e.g., AAV delivery), where the fragments are spliced together by a split intein system, thereby reconstituting a functional base editing system in the cell.

An adenovirus comprising a sequence of formula (I) 5′ITR-B.sub.1-B.sub.A-B.sub.2-B.sub.X-B.sub.B-B.sub.Y-B.sub.3-3′ITR wherein B.sub.Y comprises a transgene cassette containing four transgenes, said genes encoding a FAP-Bispecific T cell activator, CXL10, CXL9, and IFN. The disclosure also extends to a pharmaceutical composition comprising the virus, and use of the virus or formulation in treatment.

The present disclosure provides methods and compositions for treating cancers using modified immune cells that express a ROR1-specific binding protein according to certain treatment protocols including, for example, dosing regimens, infusion schedules, and patient selection criteria. In certain embodiments, presently disclosed methods and compositions are useful for treating solid cancers and/or hematological malignancies (e.g., triple-negative breast cancer (TBNC), non-small cell lung cancer (NSCLC), mantle cell lymphoma (MCL), acute lymphoblastic leukemia (ALL), or chronic lymphocytic leukemia (CLL)), wherein the methods comprise administering modified T cells that target a ROR1 antigen.

Methods of treating cancer in Non-Responder Patients with an engineered NeoTCR Product are described herein.

Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling the activity of proteins comprising a Cas9 variant fused to a functional effector domain, such as a nuclease, nickase, recombinase, deaminase, transcriptional activator, transcriptional repressor, or epigenetic modifying domain. For example, the inventive proteins provided comprise a ligand-dependent intein, the presence of which inhibits one or more activities of the protein (e.g., gRNA binding, enzymatic activity, target DNA binding). The binding of a ligand to the intein results in self-excision of the intein, restoring the activity of the protein.

Provided here in are methods and compositions for delivering a payload molecule to a charged avascular tissue or charged partially vascularized tissue in a subject, the method including administering to a subject in need thereof an effective amount of a carrier, wherein the carrier is a cationic carrier and/or has a surface charge distribution such that about 40% or greater of a continuous region of the surface is positively charged, and wherein the carrier is linked with a payload molecule, to deliver the payload molecule to the charged avascular tissue and/or the charged partially vascularized tissue.

Methods and compositions are provided for editing the genome of a cell without the use of an exogenously supplied nuclease. Aspects of the methods include contacting a cell with a targeting vector comprising nucleic acid sequence to be integrated into the target locus, where the cell is not also contacted with a nuclease. In addition, reagents, devices and kits thereof that find use in practicing the subject methods are provided.

Transgenic T cells and vectors for making transgenic T cells are described. The vectors can include a nucleic acid encoding a membrane-bound anti-IL6 (mb-alL6) single chain variable fragment (scFv), and the transgenic T cells can express mb-alL6. The transgenic T cells are useful for suppressing proliferation of IL-6-dependent cells, reducing IL-6 concentration, or both. In one embodiment, the vector is a bicistronic construct encoding the mb-alL6 and an anti-CD19-41 BB-003 chimeric antigen receptor (CAR). In another embodiment, an anti-IL-6 scFv can be linked to a 41 BB and 003 domains to form an anti-IL-6 CAR. The transgenic T cells expressing said constructs can reduce the linker risk of cytokine release syndrome (CRS) in cancer patients being treated with CAR T cell or for the treatment of autoimmune diseases and inflammatory diseases in which cytokines are involved in pathogenesis.

As disclosed herein, the assistance of a new generation of immunotoxins (split-immunotoxins) the range of cancers susceptible to elimination by oncolytic viruses and the specific toxicity of the latter can be expanded without compromising the selectivity of targeting. To accomplish this goal, a polypeptide chain of a potent bacterial toxin (including, but not limited to a catalytic subunit of a Diphtheria toxin, DTA) can be split into two benign parts, which can then be delivered to the cytoplasm of cancer cells via two independent cancer-specific pathways: (1) together with an oncolytic virus (encoded in its genome); (2) as a split-immunotoxin delivered via receptor-specific toxin entry pathways. The two parts are fused into a functional toxin only in the cytoplasm of dually targeted cancer cells by means of intein-catalyzed trans-splicing.

Provided are two split single-base gene editing systems. Intein-mediated split BE3, saKKH-BE3 and ABE7.10 are separately developed by using the existing protein structure information of spCas9 and saCas9 and splitting methods thereof, and have targeted gene mutation efficiency equivalent to that of unsplit BE3, saKKH-BE3 and ABE7.10 working system, thereby making possible package into an AAV for delivery.