Abstract: The present invention relates to cell culture media comprising inorganic ester derivatives of tyrosine and/or cysteine. The poor solubility of tyrosine and the often non-sufficient stability of cysteine in cell culture media is overcome by substituting them with an inorganic ester derivative, e.g. with a phosphorylated derivative.

Abstract: A process for producing a 3D tissue culture model by (a) printing a drop of bio-ink to a substrate; (b) printing a drop en.) of activator to the drop of bio-ink to form a hydrogel droplet; (c) repeating steps (a) and (b) in any order to form a hydrogel mold adapted to receive a drop containing cells; (d) printing a drop containing cells to the hydrogel mold; and (e) repeating steps (a) and (b) in any order to form a 3D tissue culture model comprising the cells encapsulated in the hydrogel mold.

Abstract: Provided are age-modified cells and method for making age modified cells using progerin or a progerin-like protein. The aging and/or maturation process can be accelerated and controlled for young and/or immature cells, such as a somatic cell, a stem cell, a stem cell-derived somatic cell, including an induced pluripotent stem cell-derived cell, by contacting with progerin or a progerin-like protein. Methods described by the present disclosure can produce age-appropriate cells from a somatic cell or a stem cell, such as an old cell and/or a mature cell. Such age-modified cells constitute model systems for the study of late-onset diseases and/or disorders.

Abstract: Provided is a method of producing a lacrimal gland epithelial cell from a pluripotent stem cell, a lacrimal gland epithelial cell produced by the method, and a reagent kit for inducing differentiation from a pluripotent stem cell into a lacrimal gland epithelial cell. A method of producing a lacrimal gland epithelial cell includes the following step A: Step A: a step including increasing expression of Pax6 gene in a pluripotent stem cell or transfecting PAX6 protein thereinto, and increasing expression of Foxc1 gene or Foxp1 gene in the pluripotent stem cell or transfecting FOXC1 protein or FOXP1 protein thereinto.

Abstract: A method of generating retinal pigment epithelium cells is disclosed. Cell populations comprising same and uses thereof are also disclosed.

Abstract: The present invention discloses a method of identifying agents that affect human astrocytes functionality using ex-vivo differentiated pluripotent stem cells (PSC). In addition, the use of human progenitor astrocytes or human astrocytes for the treatment of Amyotrophic Lateral Sclerosis (ALS) in a human subject is also disclosed.

Abstract: The invention provides methods of modulating follicular regulatory T (TFR) cell-mediated immune responses, follicular helper T (TFH) cell-mediated immune responses or both, and the use of those methods in the treatment of diseases or conditions mediated by TFR or TFH cells. The invention also provides novel methods for identifying TFR and TFH cells in a population of cells. The invention also provides compositions comprising TFR cells that have enhanced suppressive activity as compared wild type TFR cells. The invention also provides compositions comprising T follicular regulatory (TFR) cells isolated from the peripheral blood of a subject wherein the composition is enriched for TFR cells. Methods of making and using the compositions of the invention to modulate an immune response are also provided.

Abstract: Provided herein are T-cell receptor (TCR) fusion proteins (TFPs), T-cells engineered to express one or more TFPs, and methods of use thereof for the treatment of diseases, including cancer.

Abstract: Methods and compositions of erythroid cells that produce adult ?-hemoglobin, generated by culturing CD31+, CD31+/CD34+ or CD34+ cells from embryonic stem cells under serum-free culture conditions.

Abstract: The present invention provides methods for increasing expression of at least one co-stimulatory and/or co-inhibitory receptor on a T cell comprising contacting said T cell with an anti-CTLA4 antibody. In one aspect the co-stimulatory and/or co-inhibitory receptor is selected from the group of: PD-1, OX40, ICOS, CD137, TIM3, and LAG3. The present invention also provides methods of treating cancer in a human in need thereof comprising administering an anti-CTLA antibody and at least one additional agent directed to at least one co-stimulatory and/or co-inhibitory receptor to said human. In one aspect, the agent is directed to at least one co-stimulatory and/or co-inhibitory receptor selected from the group of: PD-1, OX40, ICOS, CD137 (4-1BB), TIM3, and LAG3.

Abstract: The present invention is related to a bone marrow-derived myeloid progenitor cell (MPC), preferably an isolated bone marrow-derived myeloid progenitor cell (MPC), wherein the bone marrow-derived myeloid progenitor cell (MPC) is expressing CD34, c-Kit (CD117) and CX3CR1.

Abstract: The present invention relates to the use of dermatopontin (DPT) or a functional fragment thereof for the maintenance of hematopoietic stem and/or progenitor cells in culture. The present invention further relates to a method for maintaining hematopoietic stem and/or progenitor cells in culture, the method comprising culturing the hematopoietic stem and/or progenitor cells in the presence of dermatopontin (DPT) or a functional fragment thereof.

Abstract: The invention provides methods of preparing a sample of viable diseased cells obtained from a human subject for clinical testing, wherein the methods inhibit anoikis and/or anoikis in the cells while maintaining the physiological functions and genomic composition of the cells when they were in vivo. In the methods of the invention, primary cells are cultured in media comprising at least one anoikis inhibitor, preferably at least one inhibitor of an intrinsic anoikis pathway and at least one inhibitor of an extrinsic anoikis pathway, under anti-anoikis atmospheric conditions, such as greater than 2% and less than 20% oxygen. Method combining multiple culturing conditions, including surface attachment under conditions that inhibit anoikis, are also provided. Compositions and kits for use in the methods of the invention are also provided.

Abstract: An in vitro co-culture system comprising cancer-associated fibroblasts (CAFs) and cancer cells for producing and maintaining cancer stem cells and uses thereof for identifying agents capable of reducing cancer cell stemness. Also disclosed herein are a paracrine network through which CAFs facilitate production and/or maintenance of cancer stem cells and the use of components of such a paracrine network for prognosis purposes and for identifying cancer patients who are likely to respond to certain treatment.

Abstract: Methods and compositions are provided for producing synthetically-derived keratin compositions and keratin products.

Abstract: The disclosure provides a cell culturing article. The cell culturing article includes a substrate having a surface, a hydrophilic copolymer layer positioned on the surface of the substrate, and a plurality of peptide chains individually conjugated to a surface of the hydrophilic copolymer layer. The hydrophilic copolymer layer is copolymerized by a plurality of polyvinyl alcohol units, a plurality of polyvinyl alcohol derivative units, and a plurality of units containing at least one carboxyl group. A method for manufacturing the cell culturing article, a method for culturing undifferentiated stem cells and a method for regulating stem cell differentiation are also provided herein.

Abstract: The present invention relates in pan to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

Abstract: An altered avian NDV that contains the coding sequence of avian interleukin-4 (IL-4), or a portion thereof, in the reverse orientation suppresses in-ovo IL-4 production via RNAi when administered to embryonic birds. An immunogenic composition containing this altered NDV is included in this invention. The altered avian NDV can, optionally contain a polynucleotide encoding a heterologous antigen from a heterologous avian pathogen and can produce said heterologous antigen in-ovo.

Abstract: The present disclosure provide viral compositions and methods for modulating adeno-associated virus properties including transduction efficiency, virus capsid assembly, viral genome packaging, capsid stability and intracellular processing. Engineered adeno-associated viruses with modifications in the N-terminal region of the capsid proteins VP1 or VP2 are provided which have varying effects on viral properties including transduction efficiency. Corresponding nucleic acids and amino acids are provided.

Abstract: A method is provided for producing an objective substance, for example, an aromatic aldehyde such as vanillin. The objective substance is produced from a carbon source or a precursor of the objective substance by using a coryneform bacterium that is able to produce the objective substance, wherein bacterium has been modified so that the activity of alcohol dehydrogenase is reduced.

Abstract: The present invention relates to beta-ketoacyl ACP synthase genes of the KASI/KASIV type and proteins encoded by these genes. The genes can be included in nucleic acid constructs, vectors or host cells. Expression of the gene products can alter the fatty acid profile of host cells. The KAS genes can be combined with a FATA or FATB thioesterase gene to create a cell that produces an increased amount of C8-C16 fatty acids. Suitable host cells include plastidic cells of plants or microalgae. Oleaginous microalga host cells with the new genes are disclosed.

Abstract: The present invention discloses a Nicotinamide phosphoribosyltransferase (nampt) mutant and use thereof. The present invention relates to a nicotinamide phosphoribosyltransferase (Nampt) mutant artificially obtained through genic site-directed mutation. An object of the present invention is to provide a Nampt mutant having a catalytic activity higher than that of a conventional wild type parent, wherein the enzymatic activity of the Nampt mutant provided in the present invention is 1.2-6.9 times of the enzymatic activity of the parent.

Abstract: The iCP-Cre recombinant protein of the present invention could mediate inactivation of a target genes, it may be used for the production of a conditional knockout mouse and be applied to investigation of the function and activation of the gene.

Abstract: A method for producing a recombinant polypeptide, comprising: (a) providing a 100 L to 25,000 L fed-batch bioreactor comprising (i) cells capable of expressing the recombinant polypeptide asfotase alfa (SEQ ID NO: 1), and (ii) a culture medium suitable for conducting such expression the culture medium comprising about 25 ?M to about 300 ?M zinc; (b) culturing the cells under conditions suitable to express the recombinant asfotase alfa wherein the pH of the culture medium is about 6.7 to about 7.1, and wherein zinc is added into said culture medium such that the zinc concentration in the culture medium is maintained at a concentration of about 25 ?M to about 300 ?M of zinc.

Abstract: The present invention relates to a Myceliophthora thermophila cell, which expresses a nucleotide sequence that codifies a recombinant beta-xylosidase enzyme comprising an amino-acid sequence having at least 70% identity with SEQ ID NO: 1, an enzymatic composition comprising said cell and/or the recombinant enzyme with beta-xylosidase activity expressed by said cell, the use of this host cell, the recombinant enzyme with beta-xylosidase activity expressed by said cell or the composition for the degradation of biomass, and a method of producing biological products, preferably bioethanol, comprising the use of said host cell, the recombinant enzyme with the beta-xylosidase activity expressed by said cell or said composition.

Abstract: Disclosed are variants of ACE2, pharmaceutical compositions comprising the variants of ACE2, and treatment methods for reducing Angiotensin II (1-8) plasma levels and/or increasing Angiotensin (1-7) plasma levels in a subject in need thereof. The disclosed variants of ACE2 may include polypeptide fragments of ACE2 having ACE2 activity for converting AngII(1-8) to Ang(1-7). Suitable subjects suitable for the disclosed methods of treatment may include subjects having or at risk for developing diabetic and non-diabetic chronic kidney disease, acute renal failure and its prevention, chronic kidney disease, severe hypertension, scleroderma and its skin, pulmonary, kidney and hypertensive complications, malignant hypertension, renovascular hypertension secondary to renal artery stenosis, idiopathic pulmonary fibrosis, liver fibrosis such as in liver cirrhosis patients, an aortic aneurysm, cardiac fibrosis and remodeling, left ventricular hypertrophy, and an acute stroke.

Abstract: The present invention provides engineered phenylalanine ammonia lyase (PAL) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered phenylalanine ammonia lyase (PAL) polypeptides. In some embodiments, the engineered PAL polypeptides are optimized to provide enhanced catalytic activity, as well as reduced sensitivity to proteolysis and increased tolerance to storage at elevated temperatures. In some embodiments, the engineered PAL polypeptides contain fewer phenylalanine residues than wild-type PAL polypeptides. The present invention also provides methods for the use of the compositions comprising the engineered PAL polypeptides for therapeutic and industrial purposes.

Abstract: The present invention concerns a novel method for the modification and/or custom-made design of artificial non-ribosomal peptide synthetases (NRPSs) from naturally available NRPSs. The artificial NRPSs are of predetermined length and amino acid composition and sequence. Via fusion of well-defined NRPS units (so-called “exchange units”) in a certain manner, using a specific sequence motif in the linker areas it is possible to construct artificial and/or modified NRPS assembly lines, which have the ability of synthesizing peptides of a desired structure.

Abstract: CRISPR/Cas Systems are provided where two or more guide RNAs having different spacer sequence lengths direct an enzymatically active Cas9 having a transcriptional regulator attached thereto to either cut a target nucleic acid or regulate expression of a target nucleic acid.

Abstract: Disclosed are processes and kits for rapid nucleic acid extraction from a nucleic acid-containing material, such as a bone, tooth or semen sample. For bone and tooth process involves providing the nucleic acid-containing material in a form suitable for nucleic acid extraction, adding a lysis buffer to the nucleic acid-containing material to obtain a mixture, mixing the mixture in a manner equivalent for about 30 seconds or longer and separating the mixture by centrifugation to obtain a liquid supernatant. The liquid supernatant contains the extracted nucleic acids which can be used for analysis including STR profiling by conventional or rapid DNA analysis. For semen the processes and kits involve applying an appropriate amount of sperm disruptive agent.

Abstract: A method of separating a target double-stranded nucleic acid molecule from a sample including the target double-stranded nucleic acid molecule and a non-target double-stranded nucleic acid molecule, including (1) mixing the sample, a pyrrole-imidazole-containing polyamide (first PI polyamide) modified with a first linker molecule and capable of specifically binding to a sequence of the target double-stranded nucleic acid molecule, and a carrier a modified with a first ligand capable of specifically binding and/or adsorbing to the first linker molecule such that a mixed solution is produced, (2) forming a complex A by binding the carrier a to the first PI polyamide with which the target double-stranded nucleic acid molecule is bound in the mixed solution, and (3) separating the complex A from the mixed solution.

Abstract: Methods of non-destructively obtaining the genotype of a plant cell from a plant sample are disclosed. The plant cell is isolated from a plant cell sample using an integrated microfluidic device. The integrated microfluidic device includes an individual cell trap located downstream from a microfluidic channel and, the microfluidic device is configured to trap the plant cell in the individual cell trap. mRNA is extracted from the plant cell by contacting the plant cell with an atomic force microscope (AFM) probe and by attracting mRNA from loci of interest to the probe end using a dielectrophoresis DEP force under the alternating current (AC) field applied to the probe. The genotype of the plant cell is determined from cDNA obtained from the extracted mRNA. Alternatively, the mRNA is analyzed to determine gene expression patterns of the plant cell.

Abstract: A dissection system has a dissection platform which has a frame, an agitation platform, a tissue section tray, a solution dispenser unit, an airflow drying unit and a waste collection unit and a specimen collector which has a tubular body, a plunger button, a hollow shaft, a piston cylinder, a motor unit, a piston, a piston spring and a piston rod. The agitation platform, the solution dispenser unit, the airflow drying unit and the waste collection unit are disposed on the frame, and the tissue section tray is removably disposed on the agitation platform. The plunger button, the hollow shaft and the piston cylinder are movably inserted in the tubular body. The piston is slidably inserted within the piston cylinder, the piston spring is biased in between the piston cylinder and the motor unit, and the piston rod is connected in between the motor unit and the piston.

Abstract: Methods are provided for assembly of a target polynucleotide sequence comprising at least a first double stranded polynucleotide (DSP) and at least second DSP, and optionally further DSPs. The method comprises an assembly reaction comprising steps including providing a first single stranded polynucleotide (SSP) comprising the polynucleotide sequence of one strand of the first DSP, and a second SSP comprising the polynucleotide sequence of one strand of the second DSP and converting the SSPs to double stranded form via a primer and polymerase-mediated extension reaction. The DSPs comprise polynucleotide sequences that are complementary to other polynucleotide sequences within the assembly reaction such that the ordering and directionality of each of the first and second, and further DSPs is determined by unique overhang pairing. Nucleic acid libraries and methods of making such libraries are also provided.

Abstract: The present invention is directed to SH3 domain derivatives having a specific binding affinity to a target molecule. In this respect, the invention provides SH3 domain derivatives of nephrocystin (NPHP1) and the Tec kinase. The invention also provides a method for the production of a library comprising recombinant derivatives of NPHP1 or the Tec kinase SH3 domains and a method for selecting from the library one or more derivatives of the SH3 domain of nephrocystin (NPHP1) or the Tec kinase having a specific binding affinity to a target molecule.

Abstract: Methods and compositions are disclosed for introducing molecular switch functionality into a protein affinity reagent to render its binding to a target molecule sensitive to an environmental trigger, such as pH, while maintaining binding affinity to the target molecule. Combinatorial libraries created by the method are also disclosed.

Abstract: Some embodiments of the present invention relate to the enrichment of non-host nucleic acids in a mixture of host and non-host nucleic acids. Some embodiments include methods for detecting pathogenic organisms from a nucleic acid sample comprising host nucleic acids and nucleic acids indicative of the pathogenic organism.

Abstract: Compositions and methods for tracking combinations of mutations in populations by coupling DNA assembly and paired-end sequencing technology with high-throughput sequencing. This allows for an unlimited number of mutation sites in a population of microorganisms to be identified, and allows sites to be sequenced across several replicates. Unique identifiers (DNA bar-codes) can be used to measure fitness data by sequencing of the barcodes multiplexed with the mutation sites with high-throughput short sequencing read technology, allowing selections to be rapidly performed on populations of known combinatorial genotypes.

Abstract: Provided herein are methods and composition for trackable genetic variant libraries. Further provided herein are methods and compositions for recursive engineering. Further provided herein are methods and compositions for multiplex engineering. Further provided herein are methods and compositions for enriching for editing and trackable engineered sequences and cells using nucleic acid-guided nucleases.

Abstract: Provided herein are methods and composition for trackable genetic variant libraries. Further provided herein are methods and compositions for recursive engineering. Further provided herein are methods and compositions for multiplex engineering. Further provided herein are methods and compositions for enriching for editing and trackable engineered sequences and cells using nucleic acid-guided nucleases.

Abstract: Hybrid reverse transcriptases formed from portions of FLVRT and MLVRT are provided.

Abstract: Inflammatory diseases in the CNS can be treated or alleviated by the administration of an oligonucleotide in an amount sufficient to reduce the influx of mononuclear cells to the central nervous system by down-regulating the expression of at least one cell surface marker. For example multiple sclerosis can be treated or at least alleviated, by the administration of an oligonucleotide in a dose effective to inhibit or reduce the influx of mononuclear and/or autoaggressive cells to the central nervous system. The oligonucleotide can be used alone, or in combination with other treatment strategies.

Abstract: Provided herein are chemically ligated guide RNA oligonucleotides (lgRNA) which comprise two functional RNA modules (crgRNA and tracrgRNA) joined by non-nucleotide chemical linkers (nNt-linker), their complexes with CRISPR-Cas9, preparation methods of Cas9-lgRNA complexes, and their uses for prevention and treatments of HIV infections in humans. Also disclosed are processes and methods for preparation of these compounds.

Abstract: Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of fibrosis. Disclosed herein are methods for decreasing A1AT mRNA and protein expression and treating, ameliorating, preventing, slowing progression, or stopping progression of liver disease, such as, A1ATD associated liver disease, and pulmonary disease, such as, A1ATD associated pulmonary disease in an individual in need thereof. Methods for inhibiting A1AT mRNA and protein expression can also be used as a prophylactic treatment to prevent individuals at risk for developing a liver disease, such as, A1ATD associated liver disease and pulmonary disease, such as, A1ATD associated pulmonary disease.

Abstract: The present invention provides methods for increasing the sensitivity of a tumour to immunotherapy, chemotherapy or radiotherapy by administering an effective amount of an oligonucleotide that inhibits the binding of miR-27a, or a variant thereof, to its target mRNA. Oligonucleotides used in the invention are typically in the form of a blockmirs used as an adjunctive therapy to inhibit tumour growth, normalise and/or improve function of tumour vasculature, and/or promote immune cell infiltration of tumours.

Abstract: The disclosed subject matter relates to brush polymer-oligonucleotide conjugates comprising oligonucleotides covalently attached to the backbone of a non-cationic, sterically congested brush polymer and the use of such polymer-oligonucleotide conjugates in antisense gene regulation and as diagnostic agents.

Abstract: The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of Atonal homolog 1 (ATOH1), in particular, by targeting natural antisense polynucleotides of Atonal homolog 1 (ATOH1). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of ATOH1.

Abstract: The present invention provides a screening method for pain suppressors, which method is characterized by using netrin-4 and/or a netrin-4 receptor to select a substance capable of inhibiting downstream signaling from netrin-4. According to the screening method of the present invention, pain suppressors useful as a preventive or therapeutic medicine for pain can be identified. The present invention also provides a pharmaceutical composition for prevention or treatment of pain, which composition comprises, as an active ingredient, a substance capable of inhibiting downstream signaling from netrin-4.

Abstract: RNA interference (RNAi) triggers and RNAi trigger conjugates for inhibiting the expression of Hif2? (EPAS1) gene are described. Pharmaceutical compositions comprising one or more Hif2? RNAi triggers optionally with one or more additional therapeutics are also described. Delivery of the described Hif2? RNAi triggers to tumor cells in vivo provides for inhibition of Hif2? gene expression and treatment of cancer.

Abstract: Compositions and methods for modifying genetic material are provided. One embodiment provides aptamers capable of binding to a site-specific DNA binding moiety to facilitate the exchange of homologous genetic information between a donor molecule and the desired target locus (aptamer-guided gene targeting or AGT). One embodiment provides an oligonucleotide containing a aptamer, preferably a DNA aptamer at the 5? end. The oligonucleotide also contains a region of homology, also referred to as donor DNA, to a desired nucleic acid, locus, or gene. The DNA binding moiety can be a nucleic acid, a protein, or a complex of proteins. In a preferred embodiment the DNA binding moiety is a homing endonuclease that cuts DNA to facilitate the modification of the DNA by the donor DNA.