Abstract: The present invention relates to methods for reducing the heterogeneity of a population of recombinant proteins produced in cell culture, said methods comprising growing host cells producing a recombinant protein in a cell culture medium wherein the cell culture medium comprises one or more cysteine/cystine analogs.

Abstract: A mold for producing a hydrogel support for a 3-dimensional (3D) cell culture, includes: an upper mold comprising a base and a plurality of upper unit molds protruding downward from the base to form accommodating portions of a female type corresponding to shapes of hydrogel supports, having through holes at bottoms thereof, and patterned at a lower portion of the base; a lower mold where a plurality of lower unit molds are patterned, the plurality of lower unit molds formed in a female type to respectively accommodate the plurality of upper unit molds protruding downward and having sealed lower portions; and an ejecting unit for separating, from the accommodating portions of the plurality of upper unit molds, the hydrogel supports that are coagulated after being inserted into the through holes respectively formed in the plurality of upper unit molds.

Abstract: Culture media, which contain albumin carrying a reduced amount of fatty acid, are useful for culturing stem cells.

Abstract: Provided herein are methods of producing, compositions comprising and uses of oligodendrogenic neural progenitor cells (o-NPCs), made using a combination of PDGFR agonist and thyroxin or a thyroxin analogue. The method includes; obtaining ventralized neural progenitor cells (NPCs), the ventralized NPCs expressing Sox2, Nkx6-1, decreased level of Pax6 compared to unpatterned NPCs, and elevated expression of HoxA4 compared to unpatterned NPCs; culturing the ventralized NPCs for about 12 to about 16 days (days 26-40 of FIG. 7; days 12 to 27 of FIG. 10) in neural expansion media (NEM) supplemented with i) PDGFR agonist for the about 12 to about 16 days and ii) thyroxine or a thyroxine analogue for the latter about 7 to about 9 days, to produce o-NPC expressing Sox2 and Nkx2.2, decreased level of Pax6 and Nkx6.1 compared to ventralized NPCs and elevated level of HoxA4 and Olig2 compared to ventralized NPCs.

Abstract: The present invention relates to an artificial antigen-presenting cell prepared from an HLA-null cell line by using a multiplex CRISPR-Cas9 system and the use thereof and, more particularly, to a novel artificial antigen-presenting cell which includes the ability to present antigens of HLA class I and a co-stimulatory molecule group transferred from an HLA-A, -B, -C null cell line generated using a multiplex CRISPR-Cas9 system and to stimulate T cells, an immunotherapeutic agent using the same, and the use thereof for treating tumors, pathogenic infections, and autoimmune diseases.

Abstract: This disclosure relates to compositions and methods of reversing senescence in T cells by interrupting vasoactive intestinal peptide (VIP) signaling and/or inhibiting phosphatidylinositol-3-kinase (PI3 kinase) inhibitor signaling and uses in managing cancer and chronic viral infections. In certain embodiments, the disclosure contemplates methods of reversing T cell senescence by mixing T cell in vitro with an agent that prevents VIP from interacting with VIP receptors and/or a PI3 Kinase inhibitor. In certain embodiments, the disclosure contemplates the expansion of senescent T cells by mixing with a PI3 kinase inhibitor, an agent that block VIP and VIP receptor signaling, a VIP degrading enzyme, and combinations thereof.

Abstract: The present invention provides a method for inducing osteogenic differentiation, the method comprising the following steps of: (1) culturing pluripotent stem cells under feeder-free conditions, (2) culturing the cells in a mixed culture medium of an osteogenic induction medium and a pluripotent stem cell medium, the mixed culture medium containing a ROCK inhibitor and a retinoic acid receptor ? or ? agonist, and (3) culturing the cells in an osteogenic induction medium containing the retinoic acid receptor ? or ? agonist. The method for inducing osteogenic differentiation according to the present invention is a simple, short-term, highly efficient and highly reproducible one-procedure method for inducing osteogenic differentiation, wherein the method is suitable for bone regeneration therapies, the development of bone metabolic drugs and the development of novel therapies for bone diseases.

Abstract: Disclosed herein are methods of producing chondrocytes from pluripotent stem cells. The invention further provides methods of regenerating cartilaginous tissue.

Abstract: Disclosed herein are methods for generating satellite cells and compositions including satellite cells.

Abstract: An object of the present invention is to prepare a functional intestinal organoid from pluripotent stem cells. An intestinal organoid is prepared from pluripotent stem cells, by the following steps (1) to (4): (1) differentiating pluripotent stem cells into endoderm-like cells; (2) differentiating the endoderm-like cells obtained in step (1) into intestinal stem cell-like cells; (3) culturing the intestinal stem cell-like cells obtained in step (2) to form spheroids; and (4) differentiating the spheroids formed in step (3) to form an intestinal organoid, the step including culture in the presence of a MEK1/2 inhibitor, a DNA methylation inhibitor, a TGF-? receptor inhibitor, and a ?-secretase inhibitor, in addition to an epidermal growth factor, a BMP inhibitor, and a Wnt signal activator.

Abstract: Provided herein are method to increase the efficiency of interspecies chimera generation.

Abstract: An in vitro model of an in vivo gastrointestinal tract including an in vitro model of an in vivo small intestine including a plurality of fermentation vessels and an in vitro model of an in vivo large intestine including a plurality of fermentation vessels is provided. A method of simulating a biotransformation of food product through the human digestive tract using an in vitro model of an in vivo gastrointestinal tract is provided.

Abstract: This disclosure relates mRNA therapy for the treatment of ornithine transcarbamylase deficiency (OTCD). mRNAs for use in the invention, when administered in vivo, encode human ornithine transcarbamylase (OTC), isoforms thereof, functional fragments thereof, and fusion proteins comprising OTC. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to effect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of OTC expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic ammonia associated with deficient OTC activity in subjects.

Abstract: Bacillus agaradhaerens strain WDG185 expresses an inulosucrase that efficiently synthesizes a broad range of IOS with a GF range of GF3-GF30. The isolated and/or purified inulosucrase, recombinantly engineered variants thereof, active fragments thereof, synthetic nucleic acids encoding the inulosucrase, its variants, or its active fragments, host cells comprising the synthetic nucleic acids, and compositions comprising the inulosucrase are provided. Methods of using the compositions include the manufacture of inulooligosaccharides.

Abstract: The present invention is directed to terminal deoxynucleotidyltransferase (TdT) variants that (i) comprise an amino acid sequence that is at least a specified percent identical to an indicated SEQ ID NOs and have at least one substitution at Q455 or at least Q455 plus at least one further substitution at G186, S248, T331, Q390, K394 or H466 (where positions are with respect to SEQ ID NO 1 and functionally equivalent positions in indicated SEQ ID NOs), (ii) are capable of template-free extension of a polynucleotide, and (iii) exhibit enhanced stability or enhanced efficiency in incorporating 3?-0-blocked nucleoside triphosphates into a polynucleotide. The invention is also directed to the use of these TdT variants for synthesizing polynucleotides of any predetermined sequence.

Abstract: The invention involves the provision of recombinant algal mutants that have a genetic modification to a nucleic acid sequence encoding a trehalose biosynthetic enzyme, and/or a genetic modification to a nucleic acid encoding an RNA binding domain. And in some embodiments either of these algal mutants can further have a genetic mutation to a nucleic acid sequence encoding an SGI1 polypeptide. Attenuation of one, two, or all three of these genes results in a mutant organism with increased lipid productivity. It was also discovered that one, two, three, or more genetic mutations can be accumulated or “stacked” in a particular mutant cell or organism to result in further increases in the production of lipid products. The lipid products of these mutants are useful as biofuels or for other specialty chemical products.

Abstract: Disclosed herein are compositions and methods for effecting alterations at a defined location in the genome of a plant cell or plant protoplast. Further disclosed are methods using an RNA-guided nuclease to alter a target nucleotide sequence in a cell or protoplast of a plant; embodiments of such methods include treatments with chemical or physical reagents or treatment of the cell or protoplast with a specific thermal regime, such as a heat treatment.

Abstract: Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting.

Abstract: The present invention relates to polypeptide having alpha-amylase activity. The present invention also relates to polynucleotides encoding the polypeptides; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the polypeptides.

Abstract: The present invention has a purpose of providing a novel ?-galactosidase enzyme useful for the production of oligosaccharides. Disclosed is a ?-galactosidase enzyme comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 4 or an amino acid sequence that is 80% or more identical to said amino acid sequence.

Abstract: A system and method for controlling metabolic enzymes or pathways in cells to produce a chemical above the levels of a wild-type strain is disclosed. The system utilizes cells, including yeasts, bacteria, and molds, having at least two genes capable of being controlled bi-directionally with light, where one gene is turned from off to on when exposed to light and another gene is turned from on to off when exposed to light, the two genes reversing when the light is turned off. Cells may utilize any number of sequences that benefit chemical production, including sequences that: encode for constitutive transcription of light-activated transcription factor fusions; encode for a metabolic enzyme; encode for a repressor; induce expression of metabolic enzymes; and an endogenous or exogenous activator expressed by a constitutive promoter, inducible promoter, or gene circuit.

Abstract: The disclosure provides methods of manufacturing products comprising allulose produced by contacting a protein having allulose 3-epimerase activity with a fructose substrate under conditions such that the fructose substrate is converted into allulose. The disclosure also provides methods of manufacturing products comprising allulose produced by providing a vector comprising a nucleic acid molecule having a polynucleotide sequence encoding a protein having allulose 3-epimerase activity.

Abstract: A method of epimerizing an (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid is provided. The method comprises contacting the (S)-1-benzylisoquinoline alkaloid with at least one enzyme. Contacting the (S)-1-benzylisoquinoline alkaloid with the at least one enzyme converts the (S)-1-benzylisoquinoline alkaloid to an (R)-1-benzylisoquinoline alkaloid.

Abstract: The present invention relates to a newly isolated bacterium belonging to the genus Microbacterium, a composition for producing psicose comprising the strain, and a method for producing psicose using the same.

Abstract: The present invention relates to a method for producing bacterially synthesized cellulose (BC) non-woven as well as to BC non-woven produced by the method and uses of such BC non-woven. The present invention also relates to an apparatus for production of the BC non-woven. Preferably, the bacterially synthesized cellulose (BC) of the present invention is biotechnologically produced nano-structured cellulose (BNC).

Abstract: This disclosure provides epimerase enzymes useful for commercial scale production of allulose from fructose. The disclosed enzymes (“epimerase variants”) are variants of Burkholderia multivorans CGD1 xylose isomerase engineered to have improved catalytic activity of about 1.5- to 2-fold compared with the parent enzyme.

Abstract: The objects of the present invention are to provide a novel PVA-degrading enzyme that the entity of which is revealed at the amino acid sequence level, a process for producing the same, a DNA encoding the enzyme, a recombinant DNA comprising the DNA, and a transformant having the recombinant DNA. The present invention solves the above objects by providing a polyvinyl alcohol-degrading enzyme having the following characteristics (1) to (3), a process for producing the same, a DNA encoding the enzyme, a recombinant DNA comprising the DNA, and a transformant having the recombinant DNA: (1) having an activity of oxidizing polyvinyl alcohol and forming hydrogen peroxide; (2) having an activity of hydrolyzing ?-diketone; and (3) exhibiting a molecular weight of 100,000±20,000 in SDS-polyacrylamide gel electrophoresis.

Abstract: Provided herein are compositions and method for producing ?-polyglutamic acid (PGA). In particular, provided herein are bacterial co-culture systems and methods for producing PGA.

Abstract: Embodiments of the invention provide multilayer analyte sensors having material layers (e.g. high-density amine layers) and/or configurations of material layers that function to enhance sensor function, as well as methods for making and using such sensors. Typical embodiments of the invention include glucose sensors used in the management of diabetes.

Abstract: A method of selecting an agent for treating a disease of a subject is disclosed which includes identifying genes which bring about resistance to a cytotoxic agent in haploid human embryonic stem (ES) cells. Once the gene is identified, the method includes analyzing the sequence and/or expression of the gene in a cell sample of the subject, wherein an alteration in the sequence and/or level of expression of the gene as compared to the sequence and/or expression of the gene in a control sample is indicative that the agent should be ruled out as a monotherapy for treating the disease in the subject.

Abstract: Devices and methods are provided to detect the presence of bacteria and small microorganisms, and to identify various microbial attributes rapidly.

Abstract: The present disclosure relates, in general, to a method for generating chimeric DNA derived from RNase L cleavage products in a sample, and detection of such RNase L cleavage products for diagnosis and treatment of inflammation and infection in a subject.

Abstract: According to one embodiment, a sensor for detecting a target substance in gas a sample includes a target substance uptake unit that brings an acetylcholine aqueous solution into contact with a gas sample to dissolve a target substance in the gas sample into the acetylcholine aqueous solution, a reaction unit that holds acetylcholinesterase and brings the solution delivered from the target substance uptake unit into contact with the acetylcholinesterase, and a detection unit that measures a change in an amount of acetylcholine decomposition product produced in the solution delivered from the reaction unit.

Abstract: The present disclosure provides a method for detecting L-serine based on cysteine desulfurase-containing living Escherichia coli cells, and belongs to the technical field of amino acid detection. The method includes the following steps: incubating an unknown sample with the cysteine desulfurase-containing living E. coli cells to produce a red substance, and qualitatively or semi-quantitatively detecting L-serine content in the unknown sample according to color changes of the red substance of the living E. coli cells, or quantitatively detecting L-serine content in the unknown sample by measuring absorbance of a lysate of the living E. coli cells. The detection method provided by the present disclosure is simple and convenient in process, few in reaction steps and stable in enzymatic activity of living cells.

Abstract: A method for sizing a DNA molecule is disclosed, which comprises the following steps of: providing a DNA sizing device, comprising: a cover substrate; a substrate disposed on the cover substrate and comprising a first hole and a second hole; and a first slit-like channel disposed between the cover substrate and the substrate, wherein two ends of the first slit-like channel respectively connects to the first hole and the second hole; loading a sample comprising a DNA molecule to the first slit-like channel through the first hole, wherein the DNA molecule moves in a direction from the first hole to the second hole; detecting and recording an intensity and an area of a distribution of the DNA molecule; and analyzing the intensity and the area to obtain the size of a DNA molecule.

Abstract: Improved methods for use in nucleic acid amplification, including multiplex amplification, where the amplification is carried out in two or more distinct phases are disclosed. The first phase amplification reaction preferably lacks one or more components required for exponential amplification. The lacking component is subsequently provided in a second, third or further phase(s) of amplification, resulting in a rapid exponential amplification reaction. The multiphase protocol results in faster and more sensitive detection and lower variability at low analyte concentrations. Compositions for carrying out the claimed methods are also disclosed.

Abstract: Provided is a nucleic acid sample-contained container including; a first nucleic acid molecule including an intended base sequence and a base sequence for detection different from the intended base sequence; and a second nucleic acid molecule free of the intended base sequence but including the base sequence for detection, wherein the nucleic acid sample-contained container includes the first nucleic acid molecule in a predetermined number. In a preferable mode, the copy number of the intended base sequence is less than 1,000, and the coefficient of variation (CV value) for the copy number is lower than 20%. In a more preferable mode, the nucleic acid molecules are artificially synthesized nucleic acid molecules. In a yet more preferable mode, the first nucleic acid molecule includes the intended base sequence in a plural number in the same molecule.

Abstract: The invention provides methods, compositions, kits and devices for the detection of target molecules. In some embodiments, the invention allows for multiplexed target molecule detection.

Abstract: The present disclosure provides compositions and methods that employ the compositions for conducting pairwise sequencing and for generating concatemer template molecules for pairwise sequencing. The concatemers can be generated using a rolling circle amplification reaction which is conducted either on-support, or conducted in-solution and then distributed onto a support. The rolling circle amplification reaction generates concatemers containing tandem copies of a sequence of interest and at least one universal adaptor sequence. An increase in the number of tandem copies in a given concatemer increases the number of sites along the concatemer for hybridizing to multiple sequencing primers which serve as multiple initiation sites for polymerase-catalyzed sequencing reactions. When the sequencing reaction employs detectably labeled nucleotides and/or detectably labeled multivalent molecules (e.g.

Abstract: Described herein are genetic recognition reagents comprising terminal aromatic moieties that bind specifically to a template nucleic acid and concatenate. Also provided are methods of using the genetic recognition reagents, e.g., to treat or diagnose a repeat expansion disorder, such as DMI.

Abstract: The present invention relates to differentiating signals of interest for target nucleic acid sequences. The present invention permits to obtain an individual signal value (i.e., variable) contained in a total signal detected at detection temperatures by using mathematical equations. The present invention based on equation-solving approach enables to obtain the individual signal value in a systematical manner, thereby providing analysis results in much more accurate and convenient manner.

Abstract: Disclosed herein, inter alia, are polynucleotides, supports, kits, and methods of use thereof for amplifying, immobilizing, and sequencing polynucleotides.

Abstract: A primer set for detecting telomerase activity, the primer set including a first primer set or a second primer set. The first primer set includes: an upstream primer selected from MTS; and a downstream primer selected from the group consisting of ACX-M4, Beacon ACX62-2C, and Beacon ACX62-10. The second primer set includes: an upstream primer selected from STS or CTS; and a downstream primer selected from the group consisting of ACX, CXT, ACX-M4, Beacon ACX62-2C, or Beacon ACX62-10. The sequences of the primers ACX, CXT, ACX-M4, Beacon ACX62-2C, Beacon ACX62-10, STS, CTS and MTS are shown as SEQ ID NOs: 1 to 8, respectively.

Abstract: In some aspects, the present disclosure provides methods for enriching amplicons, or amplification products, comprising a concatemer of at least two or more copies of a target polynucleotide. In some embodiments, a method comprises sequencing the amplicons comprising at least two or more copies of a target polynucleotide. In some embodiments, the target polynucleotides comprise sequences resulting from chromosome rearrangement, including but not limited to point mutations, single nucleotide polymorphisms, insertions, deletions, and translocations including fusion genes. In some aspects, the present disclosure provides compositions and reaction mixtures useful in the described methods.

Abstract: The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.

Abstract: A method of identifying a polynucleic acid (PNA) is presented, including the steps of providing a PNA; modifying one or more nucleobases of the PNA by addition or removal of a hydrogen bonding partner, thereby altering the base pairing capacity of the one or more nucleobases; base pairing a complementary nucleic acid to the PNA, including base pairing to at least one modified nucleobase; identifying the sequence of the complementary nucleic acid at least at the position that is complementary to at least one modified nucleobase.

Abstract: There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adapter; b) performing a single PCR cycle on the pWGAlib using a first primer (1PR) comprising from 5? to 3? a first sequencing adapter (1PR5SA) and a first primer 3? section (1PR3S) hybridizing to the reverse complementary of the WGA library universal sequence adapter; c) performing a single PCR cycle on the on the product of step b) using a second primer (2PR) comprising from 5? to 3? a second sequencing adapter (2PR5SA) different from the 1PR5SA, and a second primer 3? section (2PR3S) hybridizing to the WGA library universal sequence adapter reverse complementary; d) amplifying by PCR the product of step c) using a third primer comprising the 1PR5SA and a fourth primer comprising 2PR5SA.

Abstract: The present invention relates to methods for identifying an EoE endotype of a patient and treating the patient with one or more therapies targeted to the patient's disease endotype; and related methods for stratifying patients for clinical trials.

Abstract: CNA biomarkers can include nucleic acid sequences that are present in circulating nucleic acids obtained from circulatory systems of a population known to have cancer, but that are rarely present, if at all, in circulating nucleic acids obtained from circulatory systems of a control population. Information is received describing one or more sequences obtained from circulating nucleic acids in a population known to have cancer. Information is also received describing one or more sequences obtained from circulating nucleic acids in a control population. A cluster analysis within a predetermined portion of a genome uses the one or more sequences in the circulating nucleic acids obtained from the population known to have cancer and the one or more sequences in the circulating nucleic acids obtained from the control population. Information is generated identifying one or more biomarkers for the cancer based on a cluster search within results of the cluster analysis.

Abstract: The present invention provides diagnostic and therapeutic methods and compositions for cancer. The invention provides methods of determining whether an individual having a cancer is likely to respond to treatment comprising a pan-RAF dimer inhibitor and a MEK or PI3K inhibitor, methods of predicting responsiveness of an individual having a cancer to treatment comprising a pan-RAF dimer inhibitor and a MEK or PI3K inhibitor, methods of selecting a therapy for an individual having a cancer, and methods of treating an individual having cancer based on the presence of a biomarker of the invention (e.g., a KRAS activating mutation, e.g., a KRAS-G13D mutation, or an NRAS activating mutation).