The present invention is in the field of oncolytic viruses. The invention provides new poxviruses which are engineered to be defective for the function encoded by the M2L locus (i.e., m2 function). Such poxviruses lack a functional m2 binding activity to at least one or both of CD80 and CD86 co-stimulatory antigens. Said oncolytic poxviruses are preferably vaccinia virus having a total or partial deletion of the M2L locus. The present invention also relates to cells and compositions comprising such poxviruses and their use for treating proliferative diseases such as cancers and for preventing diseases (vaccination, especially in veterinary field). More precisely, the invention provides an alternative to the existing oncolytic viruses which are largely used in virotherapy. The m2-defective poxviruses are particularly useful for the expression of immunomodulatory polypeptides such as anti-CTLA-4 antibodies with the purposes of stimulating or improve immune response.

The present application relates to a method for reestablishing stem cells capable of forming chimeras, and cells obtained by the method. The method of the present invention is a technique for monocloning stem cells, for example, capable of forming chimeras from a heterogeneous cell population to obtain high-quality stem cells.

The invention refers to a non-therapeutic method for producing antigen-specific B cells by using the adoptive cell transfer of primed B cells, especially of spleen cells including B cells of a previously immunized non-human animal and by administering an antigen of interest to a naïve non-human animal.

Described in the present application are methods for preparing populations of hematopoietic stem cells (HSCs), e.g., autologous and/or allogenic HSCs, using mechanical stretching or Trpv4 agonisists, and methods of use of the HSCs in transplantation. In some embodiments, the methods include providing a population comprising hemogenic endothelial (HE) cells, and (i) contacting the HE cells with an amount of an agonist of transient receptor potential cation channel-subfamily vanilloid member 4 (Trpv4); and/or (ii) subjecting the cells to cyclic 2-dimensional stretching, for a time and under conditions sufficient to stimulating endothelial-to-HSC transition. Also provided herein are methods for treating subjects who have, bone marrow, metabolic, and immune diseases; the methods include administering to the subject a therapeutically effective amount of hematopoietic stem cells (HSCs) obtained by a method described herein.

Disclosed herein are chimeric mammals, such as a chimeric mammal comprising cells derived from at least a first mammal and a second mammal, wherein the cells from the first mammal comprise a genetic modification at one or more loci and the cells from the second mammal form at least one organ or tissue, wherein the first and second mammals are different species.

A method for establishing an animal model of hepatocellular carcinoma (HCC) bone metastasis, the method including: 1) establishing 97H and LM3 cell clones with stable expression of firefly luciferase (LUC); 2) allowing HCC cells to form bone metastasis in nude mice via intratibial injection; 3) reproducing HCC bone metastasis in nude mice via intracardiac injection of tumor cells; and 4) isolating a sub-population of tumor cells that targets metastasis to bone. The 97H and the LM3 are highly metastatic HCC cell lines transfected with luciferase gene. BALB/cA-nu mice are 4-5 weeks old and maintained in laminar flow cabinets under SPF conditions and received human care throughout an entire study. A cell number for intratibial injection is 0.5×10.sup.6, and a cell number for intracardiac injection is 1×10.sup.6.

The present invention provides an immuno-deficient animal useful as an animal model for a human disease associated with a first mutation of a target gene, wherein the animal comprises (a) a first human xenograft comprising the target gene comprising the first mutation; (b) a second human xenograft comprising the target gene but lacking the first mutation, wherein the first human xenograft and second human xenograft are isogenic. Also provided here are methods of producing the animal model and methods of using such animal model.

The present disclosure provides methods and compositions for enhanced editing of genomic DNA. For example, in some embodiments, a subject method is a method of editing genomic DNA of a eukaryotic cell and the method includes introducing into a eukaryotic cell a composition comprising: (a) a linearized non-homologous DNA composition and (b) a genome targeting composition (which includes a genome editing endonuclease, or a nucleic acid encoding the genome editing endonuclease). In some cases in which the genome editing endonuclease is a CRISPR/Cas endonuclease, the genome targeting composition can also include a corresponding CRISPR/Cas guide RNA.

The present invention provides a method for treating wounds by applying cells as described in this application. In one aspect the method provides treatment for cutaneous wounds. In general embodiments the cells are delivered to the wound without being attached to a functionalized substrate in the delivery vehicle.

Compositions and methods are provided for integrating one or more genes of interest into cellular DNA without substantially disrupting the expression of the gene at the locus of integration, i.e., the target locus. These compositions and methods are useful in any in vitro or in vivo application in which it is desirable to express a gene of interest in the same spatially and temporally restricted pattern as that of a gene at a target locus while maintaining the expression of the gene at the target locus, for example, to treat disease, in the production of genetically modified organisms in agriculture, in the large scale production of proteins by cells for therapeutic, diagnostic, or research purposes, in the induction of iPS cells for therapeutic, diagnostic, or research purposes, in biological research, etc. Reagents, devices and kits thereof that find use in practicing the subject methods are also provided.