Abstract: Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a cell and compositions for such methods. Methods herein comprise contacting the cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the cell.

Abstract: Provided herein are multiplex crRNAs and multiplex sgRNAs, as well as RNA molecules thereof. Also provided are compositions and kits including the multiplex crRNAs and sgRNAs, which can be used in a multiplex targeted gene activation (mTGA) system. Also provided are methods that include administering a therapeutically effective amount of the mTGA system to a subject. In some examples, the method treats a disease associated with reduced or no expression of a gene, such as type I diabetes, Duchenne muscular dystrophy, a liver disease, or acute kidney disease.

Abstract: Compositions for generating a pancreatic beta-like cell population from a population of undifferentiated cells and methods of use thereof, are provided. The method is an 8 stage process interrupted by a priming step, and it includes; using a chemically defined protocol for the efficient generation of pancreatic progenitors (PPs); improved assembly PPs into 3D clusters; a priming step which uses a chemical/factor cocktail (PP-10C) to maintain 3D-PPs status and enhances their potential to differentiate into ? cells ;and a 3-step differentiation protocol using select chemical cocktails that efficiently converts PP-10C-treated 3D-PPs into functional ? cells. The disclosed methods result in a population of functional ? cells which expresses pancreatic cell markers selected from the group of c-peptide, the transcription factors NKX6.1, PDX1, PAX6, NEUROD1 and are INS+, and which do not express substantial levels Glucagon (GCG). The functional ? cells can be used to treat conditions such as diabetes.

Abstract: Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a non-dividing cell and compositions for such methods. Methods herein comprise contacting the non-dividing cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the non-dividing cell.

Abstract: Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a non-dividing cell and compositions for such methods. Methods herein comprise contacting the non-dividing cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the non-dividing cell.

Abstract: Compositions and methods modulating the steady state of cells are provided. The compositions include metabolites (C1 metabolites and C1 metabolite cocktails (C1-MIM) for use in inducing cells into a different state from their steady state, for example, into a less differentiated state, when compared to their original state before treatment. The C1 metabolites include methionine, SAM (S-adenosyl methionine), threonine, glycine, putrescine, and cysteine. The metabolites are used to supplement cell culture media, and accordingly, cells culture media supplemented with the disclosed metabolites (MIM supplemented media) are also provided. The method includes: contacting a cell with the C1 metabolites for a sufficient period of time to result in reprograming the cell into a different state from their steady, for example, into a less differentiated state having progenitor-like characteristics (MIM-Cells).

Abstract: Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a non-dividing cell and compositions for such methods. Methods herein comprise contacting the non-dividing cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the non-dividing cell.

Abstract: Provided herein are blastoids and methods for producing the same that are obtained from an extended pluripotent stem (EPS) cell. The herein-disclosed methods provide a unique and highly malleable in vitro system for studying early preimplantation development. Also provided are EPS-blastoids derived from a somatic cell.

Abstract: Methods are provided for treating osteoarthritis by administering ?Klotho protein and sTGF?-R2 protein to a site within a mammal exhibiting symptoms of osteoarthritis, such as a knee joint. The ?Klotho protein and the sTGF?-R2 protein are both present at the osteoarthritic site.

Abstract: Provided herein are methods and compositions for editing a target genome in a cell comprising contacting the cell with (i) a single homology arm construct comprising a replacement sequence and a targeted endonuclease cleavage site; and (ii) a targeted endonuclease, wherein the replacement sequence comprises at least one nucleotide difference compared to the target genome and wherein the target genome comprises a sequence homologous to the targeted endonuclease cleavage site.

Abstract: Provided are modified guide RNAs (gRNAs), including dead guide RNAs (dgRNAs) with increased GC content and/or decreased repetitive content, as well as compositions and kits including such dgRNAs, which can be used in a targeted gene activation system, for example to increase expression of a gene to treat a disease in vivo. Such methods increase targeted gene expression, without creating DNA double strand breaks (DSBs).

Abstract: This disclosure provides ungulate embryonic stem cells (ESCs) derived from the inner cell mass of pre-implantation blastocysts or pluripotent cells from embryos. From an agricultural and biomedical perspectives, the derivation of stable ESCs from domestic ungulates is important for genomic testing and selection, genetic engineering, and providing an experimental tool for studying human diseases. Cattle are one of the most important domestic ungulates that are commonly used for food and bioreactors.

Abstract: Disclosed herein are homology-independent targeted integration methods of integrating an exogenous DNA sequence into a genome of a non-dividing cell and compositions for such methods. Methods herein comprise contacting the non-dividing cell with a composition comprising a targeting construct comprising the exogenous DNA sequence and a targeting sequence, a complementary strand oligonucleotide homologous to the targeting sequence, and a nuclease, thereby altering the genome of the non-dividing cell.

Abstract: 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.

Abstract: Provided herein are self-renewable nephron progenitor cell (srNPC) and methods for making and using.

Abstract: Methods of preventing the transmission of a mitochondrial disease, disorder, or condition using mitochondria-targeted enzymes or mRNA encoding mitochondria-targeted enzymes. The methods as described herein can specifically eliminate mitochondrial DNA (mtDNA) mutations in the germline.

Abstract: Methods for heart regeneration are provided. The invention provided herein includes methods of modulating proliferation of cardiomyocytes using small molecules and micro RNAs. In embodiments, the methods provided may be used to increase proliferation or cardiomyocytes. Further provided are methods to be used for the treatment of myocardial infarction.

Abstract: Methods for heart regeneration are provided. The invention provided herein includes methods of modulating proliferation of cardiomyocytes using small molecules and micro RNAs. In embodiments, the methods provided may be used to increase proliferation or cardiomyocytes. Further provided are methods to be used for the treatment of myocardial infarction.

Abstract: Methods for heart regeneration are provided. The invention provided herein includes methods of modulating proliferation of cardiomyocytes using small molecules and micro RNAs. In embodiments, the methods provided may be used to increase proliferation or cardiomyocytes. Further provided are methods to be used for the treatment of myocardial infarction.

Abstract: Provided herein are, inter alia, methods of generating vascular progenitor cells from primary cells without reprogramming the primary cell into a pluripotent stem cell. The vascular progenitors provided herein may be used to form endothelial cells or smooth muscle cells. Further provided are isolated mutlitpotent cells useful to form vascular progenitor cells.