Provided herein are methods and compositions for batch production of producer cells using fluorescence activated cell sorting (FACS). In some aspects, the disclosure provides a drug-selection-free method for batch production of producer cells using FACS. Such batch production methods and compositions can be further utilized to generate clonal populations of producer cells, e.g., for large-scale manufacturing of a polypeptide of interest.

Disclosed is a high-throughput transcriptional assay format in Actinomycete bacteria, and Streptomyces spp. in particular, that leverages eGFP, inserted both at a neutral site and inside the biosynthetic cluster of interest, as a read-out for secondary metabolite synthesis. Using this approach, a silent gene cluster in Streptomyces albus J1074 was induced. The cytotoxins etoposide and ivermectin were revealed as potent inducers, allowing the isolation and structural characterization of nearly 20 novel small molecule products of the chosen cluster. One of these molecules is a novel antifungal, while several others inhibit a cysteine protease implicated in cancer. Studies addressing the mechanism of induction by the two elicitors led to the identification of a pathway-specific transcriptional repressor that silences the gene cluster under normal growth conditions. The successful implementation of this approach will allow future discovery of cryptic metabolites with useful bioactivities from Actinomycete bacteria.

A platform for testing cell response to biochemical agents. The TrophoWell™ includes a well which contains a gel, and a plurality of capillaries that open into it. Cells and various biochemical agents such as drugs and growth factors are flown through those capillaries. The platform allows for the evaluation of cell response by imaging. The platform is a cost effective testing platform and can be used in the fields for drug discovery and personalized medicine.

The molecular mechanism of the function of TBC1D15 in NOTCH activation and stabilization is associated with maintenance and expansion of tumor-initiating stem-like cells (TICs). We herein demonstrate that TBC1D15 preferentially binds an NUMB isoform and NOTCH1-PEST domain and stabilizes NOTCH1 protein. This interaction enhances phosphorylation of NUMB, taking it away from NUMB-mediated ubiquitin degradation of NOTCH. Thus, TBC1D15 acts as a switch to augment NOTCH signaling in TICs. We also have identified inhibitors of TBC1D15 such as latanoprost acid and a salt thereof, which blocks the NOTCH-TBC1D15 interaction and selectively kills CD133-positive TICs, resulting in a reduction in the tumor size in a patient-derived, hepatocellular carcinoma xenograft model.

The current invention relates to an improved method for measuring modulating effects of compounds on epithelial cell barrier function. The methods allows for high throughput screening of test compounds individually or in combination. Compounds improving the epithelial barrier function or compounds negatively influencing epithelial barrier function can be analyzed with the method described herein.

Methods of screening agents in a cardiomyocyte population are provided. The cardiomyocyte population may be differentiated from a dystrophin knockout iPSC line. High-throughput methods of screening agents in a cardiomyocyte population that has been differentiated from a dystrophin knockout iPSC line are also provided. The methods may include determining an effect of the agents on membrane barrier function by using a cell viability assay. Methods of making dystrophin knockout iPSC lines, making dystrophin knockout iPSC derived cardiomyocytes, and modeling dystrophin deficient cardiomyopathy are also provided.

This disclosure relates to methods for expanding inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells) and differentiating inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells) to inner ear hair cells (e.g., atonal homolog 1 (Atoh1)+ inner ear hair cells) and the use of the inner hear supporting cells and hair cells, e.g., for identifying candidate therapeutic compounds for the treatment of hearing loss and balance loss. Additionally, the methods described herein can be used in the treatment of a subject having hearing loss and balance loss that would benefit from increased proliferation and differentiation of inner ear supporting cells (e.g., Lgr5+ inner ear supporting cells).

The present invention relates to a melanogenesis detection method using FAM86A, and the like. The level of FAM86A of the present invention decreases according to an increase in the amount of melanin secretion or formation, and thus the present invention can whiten the skin by using protein FAM86A or an agonist thereof, and can prevent, treat or alleviate melanin-deficiency diseases such as vitiligo since the formation and secretion of melanin is promoted when FAM86A is inhibited. Therefore, the present invention is expected to be used in various ways, such as a composition for skin whitening using protein FAM86A or an agonist thereof, and as a composition for preventing and treating melanin deficiency diseases including vitiligo and canities by using a FAM86A inhibitor.

A recombination expression vector can be used for screening cartilage disease treatment agents. A cell line is transformed using the expression vector. A method using the foregoing screen drugs that are effective in treating cartilage diseases, and since all constituent factors are composed of human-derived genetic factors, new drugs selected through this system are considered to be more effective in treating human cartilage diseases. Furthermore, using additional transformation, it can be evaluated whether genes having unknown functions can be used to treat cartilage diseases. This establishes the ability to not only compare the cartilage disease treatment functions of various drugs, but also to evaluate the optimal treatment concentration and indirect cytotoxicity. Moreover, 2-anthraquinonecarboxylic acid, which is a novel substance having cartilage regeneration efficacy discovered through the screening system, is utilizable in the treatment of various cartilage disease.

The present invention provides a production method of a retinal pigment epithelial cell containing the following steps: (1) a first step for culturing a pluripotent stem cell in a medium containing at least one kind selected from the group consisting of an FGF receptor inhibitor and an MEK inhibitor for a period of not more than 30 days, and (2) a second step for culturing the cell obtained in the first step in a medium containing at least one kind selected from the group consisting of a Rho signal transduction pathway inhibitor and an apoptosis inhibitor to form a retinal pigment epithelial cell. The method can produce retinal pigment epithelial cells from pluripotent stem cells more efficiently and conveniently.