Abstract: In certain embodiments an optimized derivative of the CCLc-?AS3-FB lentiviral vector termed (CCLc-mGata/ANK-CoreLCR-?AS3-FB), is provided which is capable of driving lineage-restricted expression of a beta-globin gene (e.g., an anti-sickling ?-globin like gene (?A83)). In certain embodiments the vectors described herein comprise novel defined LCR HS core sequences (HS2(˜420 bp), HS3?40 bp), HS4(˜410 bp)) which can be used to replace the putative LCR HS sequences present within the “mini-LCR” (˜3.6 kb reduced to ˜1.2 kb) to produce an “optimized mini-LCR”.

Abstract: The present invention provides means and methods for the generation of midbrain organoids which are useful for studying neurodevelopmental and neurodegenerative diseases. Neuroepithelial stem cells serve as a starting population for the generation of midbrain organoids by contacting them with differentiation medium under agitating conditions in three-dimensional cell culture comprising a matrix.

Abstract: The purpose of the present invention is to provide an agent for restoring a visual function or an agent for preventing the deterioration in a visual function, which has an excellent visual function restoring ability. The agent for regenerating a visual function or the agent for preventing the deterioration in a visual function according to the present invention contains, as an active ingredient, a chimeric protein having both an amino acid sequence for a microorganism-origin ion-transporting receptor rhodopsin and an amino acid sequence for an animal-origin G-protein-coupled receptor rhodopsin. The chimeric protein is preferably one in which an amino acid sequence for a cytoplasm-side second loop and/or a cytoplasm-side third loop in the amino acid sequence for the microorganism-origin ion-transporting receptor rhodopsin is substituted by an amino acid sequence for a cytoplasm-side second loop and/or a cytoplasm-side third loop in the G-protein-coupled receptor rhodopsin.

Abstract: Methods of preventing or treating rheumatoid arthritis (RA) in a subject by introducing the DRB1*04:01K71E mutation that is resistant to RA. The resistant allele is introduced into the subject having or at risk of developing RA, using a HLA CRISPR/Cas9 vector that targets codon 71 in the HLA allele DRB1*04:01, introducing a single A to G point mutation in codon 71 by homology directed repair to alter the lysine at position 71 of the expressed protein to glutamic acid. This modified allele is affected in the subject's hematopoietic stem cells, which are then expanded and transplanted back into the patient. This microgene therapy confers RA-resistance via an autologous transplant. The invention includes isolated nucleic acids, vectors, recombinant viruses, cells, and pharmaceutical compositions to modify the HLA DRB1*04:01 allele.

Abstract: The present disclosure provides CasX proteins, nucleic acids encoding the CasX proteins, and modified host cells comprising the CasX proteins and/or nucleic acids encoding same. CasX proteins are useful in a variety of applications, which are provided. The present disclosure provides CasX guide RNAs that bind to and provide sequence specificity to the CasX proteins, nucleic acids encoding the CasX guide RNAs, and modified host cells comprising the CasX guide RNAs and/or nucleic acids encoding same. CasX guide RNAs are useful in a variety of applications, which are provided. The present disclosure provides archaeal Cas9 polypeptides and nucleic acids encoding same, as well as their associated archaeal Cas9 guide RNAs and nucleic acids encoding same.

Abstract: Provided is a method of preparing a somatic cell including converting a differentiated somatic cell of a mammal to other somatic cell by culturing the differentiated somatic cell in a medium for inducing differentiation of the somatic cell other than the differentiated somatic cell in the presence of a TGF-? pathway inhibitor.

Abstract: A method for treating a patient suffering from a neuronal hypo-kinetic disease or a neuronal hyper-kinetic disease by modulating neuronal activity in the: internal globus pallidus (GPi), in the anterior motor thalamus and/or in the external globus pallidum (GPe) and/or in the subthalamic nucleus (STN) by utilizing suppressor and/or enhancer DREADDs is provided.

Abstract: Disclosed herein are high transducing replication defective herpes simplex virus (HSV) vectors of McKrae strain.

Abstract: This invention relates to a transgenic vector for transducing cells of a mammal's CNS and transgenically expressing a protein in the mammal's CNS. The transgenic vector comprises a virus-derived vector, a nucleic acid sequence encoding the protein, and an endogenous ATP1A3 promoter sequence. This invention also relates to a composition comprising a recombinant AAV vector comprising a nucleic acid sequence encoding a ATP1A3 protein, in a form compatible with administration into the CNS. The invention also relates to a method for treating a subject having a neurological disorder associated with mutations in the ATP1A3 gene and a method for delivering a transgenic ATP1A3 DNA to the central nervous system of a mammal by administering the recombinant AAV vector into the mammal's CNS.

Abstract: An object of the present invention is to provide novel mesenchymal stem cells demonstrating superior therapeutic effects against various diseases, a novel pharmaceutical composition containing these mesenchymal stem cells, and a method for preparing the same. The present invention relates to ROR1-positive mesenchymal stem cells. The ROR1-positive mesenchymal stem cells are preferably positive for CD29, CD73, CD90, CD105 and CD166 and are derived from umbilical cord or adipose tissue.

Abstract: A method of manufacturing a multilayered cell sheet of neural crest stem cells (NCSCs), includes: (1) isolating and culturing NCSCs from peripheral nerves; (2) embedding the cultured NCSCs in a hydrogel; (3) culturing the hydrogel comprising the NCSCs embedded therein under stressed culture conditions in which a physical support is applied; and (4) culturing the resulting hydrogel of step (3) under non-stressed culture conditions in which a physical support is removed.

Abstract: Disclosed herein are methods for generating universal MHC/HLA-compatible hematopoietic progenitor cells and methods for generating custom patient-specific MHC/HLA-compatible hematopoietic progenitor cells. Compositions comprising the universal and custom hematopoietic progenitor cells and therapeutic applications thereof are also disclosed.

Abstract: The disclosure relates to methods and compositions for regulating expression of DUX4. Specifically, the disclosure provides a recombinant gene editing complex comprising: a recombinant gene editing protein; and, a nucleic acid encoding a guide RNA (gRNA) that specifically hybridizes to a target nucleic acid sequence encoding a D4Z4 macrosatellite repeat region, wherein binding of the complex to the target nucleic acid sequence results in inhibition of DUX4 gene expression. In some aspects, methods described by the disclosure are useful for treating a disease associated with aberrant DUX4 expression (e.g., facioscapulohumeral muscular dystrophy, FSHD).

Abstract: The present invention relates, in general, to Pompe disease and, in particular, to a methods of treating Pompe disease and to compounds/constructs suitable for use in such methods.

Abstract: Provided is a cell culture method comprising the step of culturing cells using a medium containing a laminin fragment having integrin binding activity, the method not comprising the step of coating a culture vessel with a laminin or a laminin fragment before seeding the cells in the culture vessel. The cell culture method of the present invention uses a smaller amount of a laminin fragment and still achieves a comparable culture efficiency as compared with the conventional cell culture method that uses a culture vessel precoated with a laminin or a laminin fragment.

Abstract: This invention relates to polynucleotides comprising a nucleotide sequence encoding a PPT1 polypeptide or a fragment thereof, vectors (viral or non-viral vectors) comprising the same, and methods of using the same for delivery of the open reading frame to a cell or a subject and to treat infantile neuronal lipofuscinosis (infantile Batten disease). The polynucleotides comprise an optimized CLN1 open reading frame.

Abstract: The present disclosure relates to compositions and methods for culturing a population of hepatocytes in vitro, comprising co-culturing the population of hepatocytes with at least one non-parenchymal cell population and incubating the co-culture in culture medium, wherein the co-culture is periodically incubated in culture medium that does not comprise serum (serum-free culture medium).

Abstract: A method of preparing a ddx27-deletion zebrafish mutant, including: determining a target of ddx27 knockout on a sixth exon of the ddx27 in a zebrafish and designing a gRNA sequence; using primers T7-ddx27-sfd and tracr rev for PCR amplification with a pUC19-gRNA scaffold plasmid as a template; purifying and transcribing the PCR product obtained in vitro to produce gRNA; introducing the gRNA and a Cas9 protein into the zebrafish; and culturing the zebrafish to obtain a zebrafish ddx27 mutant of stable inheritance. In addition, the application also discloses a phenotype of the ddx27-deletion zebrafish mutant, which plays an important role in investigating the biological function.

Abstract: A method of growing primary human epithelial cells, in particular human epithelial cells using a basal formula containing individual (a) amino acids, (b) vitamins, (c) trace elements, and (d) other organics such as linoleic acid. The basal medium may be a mixture of amino acids, vitamins, and salts that constitute the basic media that is used to culture epithelial cells over a number of population doublings, e.g., over at least one week, while maintaining a normal phenotype and exerting low stress on the cultured cells, and maintaining lineage heterogeneity.

Abstract: Provided is a method for inducing a skeletal muscle cell including a step of introducing MyoD family gene or an expression product thereof and Myc family gene or an expression product thereof into a somatic cell of a mammal.