Methods are disclosed herein for producing human hepatocytes from human induced pluripotent stem cells. Also provided are transgenic rats for the expansion of human hepatocytes, such as those produced using the methods disclosed herein.

Provided are a three-dimensional pancreatic organoid derived from the pancreas of a genetically modified mouse, a method for fabricating the three-dimensional pancreatic organoid, and use of the three-dimensional pancreatic organoid for drug effect verification and/or drug screening.

Disclosed herein are methods for producing genetically modified cells expressing HLA-G (e.g., cell surface HLA-G) persistently, and nucleic acid compositions useful for generating such genetically modified cells. Also disclosed are cell therapy methods that utilize genetically modified cells that express HLA-G persistently. The HLA-G genetic modifications described herein provide the cells with characteristics of reduced immunogenicity and/or improved immunosuppression, such that these cells have the promise of being universal or improved donor cells for transplants, cellular and tissue regeneration or reconstruction, and other therapies.

A method of modulating some or all copies of a gene in a cell is provided including introducing into a cell one or more ribonucleic acid (RNA) sequences that comprise a portion that is complementary to all or a portion of each of the one or more target nucleic acid sequences, and a nucleic acid sequence that encodes a Cas protein and maintaining the cells under conditions in which the Cas protein is expressed and the Cas protein binds and modulates the one or more target nucleic acid sequences in the cell.

Genetically modified cells, tissues, and organs for treating or preventing diseases are disclosed. Also disclosed are methods of making the genetically modified cells and non-human animals.

We disclose that a target organ such as kidney can be regenerated by complementing a developmental deficiency leading to a lack of development of the target organ in a non-human first mammal by injecting a somatic cell nuclear transfer cell (SCNT cell) into a developed blastocyst of the non-human first mammal. We also disclose a method for producing a target organ, using an SCNT cell, in a living body of a non-human first mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a second mammal that is an individual different from the non-human first mammal.

The present invention provides a kidney production method including a step of tissue-specifically removing a metanephric mesenchyme of a metanephros of a non-human animal; a step of transplanting a human kidney precursor cell into the metanephros; and a step of advancing development of the metanephros, which is a step in which the transplanted human kidney precursor cell is differentiated and matured to form a part of the kidney.

The present invention provides an organ production method including a step of tissue-specifically removing a first portion of an organ of a non-human animal partway through development thereof; a step of transplanting, into a region from which the first portion has been removed, an organ precursor cell which is allogeneic or xenogeneic to the non-human animal; and a step of advancing development of the organ, which is a step in which the transplanted organ precursor cell is differentiated and matured to form a part of the organ.

Human or humanized tissues and organs suitable for transplant are disclosed herein. Gene editing of a host animal provides a niche for complementation of the missing genetic information by donor stem cells. Editing of a host genome to knock out or debilitate genes responsible for the growth and/or differentiation of a target organ and injecting that animal at an embryo stage with donor stem cells to complement the missing genetic information for the growth and development of the organ. The result is a chimeric animal in which the complemented tissue (human/humanized organ) matches the genotype and phenotype of the donor. Such organs may be made in a single generation and the stem cell may be taken or generated from the patient's own body. As disclosed herein, it is possible to do so by simultaneously editing multiple genes in a cell or embryo creating a niche for the complemented tissue. Multiple genes can be targeted for editing using targeted nucleases and homology directed repair (HDR) templates in vertebrate cells or embryos.

Genetically modified cells, tissues, and organs for treating or preventing diseases are disclosed. Also disclosed are methods of making the genetically modified cells and non-human animals.