Abstract: Cranial placodes are embryonic structures essential for sensory and endocrine organ development. The efficient derivation of cranial placodes from human pluripotent stem cells is disclosed where the timed removal of the BMP inhibitor Noggin, a component of the dual-SMAD inhibition strategy of neural induction, triggers placode induction at the expense of CNS fates. Further fate specification at the pre-placode stage enables the selective generation of placode-derived trigeminal ganglia capable of in vivo engraftment, mature lens fibers and anterior pituitary hormone-producing cells that upon transplantation produce hormones including, but not limited to, human growth hormone and adrenocortiocotropic hormone in vivo. Alternatively, anterior pituitary hormone-producing cells are generated in cell culture systems in vitro.

Abstract: A method of creating skin tissue is described, particularly, an in vitro or ex vivo method for creating skin tissue. The invention extends to the use of agents that disrupt the LINC complex in a skin cell to create the skin tissue, and to using the created tissue in an assay to identify or screen anti-ageing compounds. The invention further extends to model skin tissues per se, uses thereof and to kits for creating such model skin tissues.

Abstract: Methods are provided for the cell-based delivery of collagen VII for the treatment of Epidermolysis Bullosa and corneal erosion. The disclosure also provides a composition and a pharmaceutical composition comprises, comprise, or alternatively consist essentially of, or yet further consist of a keratinocyte sheet or a corneal cell sheet.

Abstract: The invention is directed to modified T cells, methods of making and using isolated, modified T cells, and methods of using these isolated, modified T cells to address diseases and disorders. In one embodiment, this invention broadly relates to TCR-deficient T cells, isolated populations thereof, and compositions comprising the same. In another embodiment of the invention, these TCR-deficient T cells are designed to express a functional non-TCR receptor. The invention also pertains to methods of making said TCR-deficient T cells, and methods of reducing or ameliorating, or preventing or treating, diseases and disorders using said TCR-deficient T cells, populations thereof, or compositions comprising the same.

Abstract: The invention is directed to modified T cells, methods of making and using isolated, modified T cells, and methods of using these isolated, modified T cells to address diseases and disorders. In one embodiment, this invention broadly relates to TCR-deficient T cells, isolated populations thereof, and compositions comprising the same. In another embodiment of the invention, these TCR-deficient T cells are designed to express a functional non-TCR receptor. The invention also pertains to methods of making said TCR-deficient T cells, and methods of reducing or ameliorating, or preventing or treating, diseases and disorders using said TCR-deficient T cells, populations thereof, or compositions comprising the same.

Abstract: The present invention provides methods for isolating and cryopreserving tumor infiltrating lymphocytes (TILs) and producing therapeutic populations of TILs, including methods via use of a kit and a semi-automatic device for aseptic disaggregation, enrichment, and cryopreservation of a resected tumor prior to expansion of the TIL population. The present invention also provides methods for expansion, and/or stabilization of TILs, for instance UTILs, compositions involving the same and methods of treatment involving the same.

Abstract: Several methods are described for generating mycelial scaffolds for use several technologies. In one embodiment, a mycelial scaffold is generated using a perfusion bioreactor system for cell-based meat technologies. In another embodiment, a mycelial scaffold is prepared for biomedical applications. The mycelial scaffolds may be generated from a liquid medium or from a solid substrate.

Abstract: Methods and compositions for generating human pancreatic beta cells, and the progenitor cells thereof, from human pluripotent stem cells using chemically defined culture media are disclosed. The methods allow for the initial generation of FOXA2+HNF1b+ dorsal foregut endodermal cells (DFECs), which can be further differentiated to human PTF1A+PDX1+ pancreatic progenitor cells (PPCs) and human INS+PDX+ pancreatic beta cells (PBCs) according to the methods provided. Culture media, isolated cell populations and kits are also provided.

Abstract: Methods for generating CD31+CD34+CD143+CD309+GATA2+FLI1+RUNX1+vWF+hemogenic endothelial cells (HECs) are provided using chemically-defined culture media that allow for generating HECs from early mesoderm progenitors in as little as three days and from pluripotent stem cells in as little as five days. Culture media, isolated cell populations and kits are also provided.

Abstract: Mechanical chopping has been successfully used to expand both fetal and iPSC-derived neural progenitor cells to scales suitable for early phase clinical trials. However, this method is time-consuming, labor-intensive, and challenging to implement at larger scales. Described herein are methods, apparatuses and systems for a novel in-line passaging technique that maintains the expansion rate and cellular identity of mechanical chopping but that is faster, scalable, and can be implemented in a fully sealed system.

Abstract: Disclosed herein are modular viral genomes comprising multiple individual DNA fragments, where each DNA fragment comprises a nucleic acid and at least one viral adaptor region at a terminal end of the nucleic acid. Individual DNA fragments may be assembled based on the homologous viral adaptor regions of each DNA fragment. Also disclosed herein are methods of producing a modular viral genome.

Abstract: Disclosed herein are compositions comprising exogenous antimicrobial agents and methods of use.

Abstract: The present invention relates to an oncolytic virus and the use thereof, specifically, an oncolytic virus having suppressed thymidine kinase (TK) gene expression and comprising genes encoding granulocyte-macrophage colony-stimulating factor (GM-CSF) and a complement regulatory protein; and the use of such an oncolytic virus. The oncolytic virus of the present invention maintains its efficacy even when administered intravenously, and thus, it may also be applied to the treatment of various solid cancers and metastatic cancers in addition to superficial solid cancers. In addition, the oncolytic virus of the present invention acquires resistance to complement attack by expressing a complement regulatory protein on the surface of the virus, and thus, it is stable in the blood, and it maintains stable oncolytic activity when intravenous injection, and thus, it may reduce effective viral dosage to minimize the side effects of anti-cancer drugs.

Abstract: Human milk oligosaccharides (HMOs) may be used e.g. as functional ingredients in infant nutrition, medical nutrition, functional foods and animal feed. There is still a need of improved means of producing HMOs. The present invention provides genetically modified microorganisms for the improved production of HMOs and HMO production methods using the same. The microorganisms of the invention may have one or more yield-enhancing modifications, including an inducible lysis system, which allows for the easy extraction of intracellular and extracellular HMOs, lactose permease mutants, which may increase intracellular lactose levels, or chaperones, which may increase intracellular availability of key enzymes for the production of HMOs.

Abstract: The present invention relates a variant polypeptide having geranylgeranyl pyrophosphate synthase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a geranylgeranyl pyrophosphate synthase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids at positions 92, 100 or 235 said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having geranylgeranyl pyrophosphate synthase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: Disclosed herein, are prime editors for prime editing. Also disclosed are engineered reverse transcriptases and methods of using same for prime editing.

Abstract: The present disclosure provides compositions, methods, kits, systems and apparatus that are useful for nucleic acid polymerization. In particular, modified polymerases and biologically active fragments thereof are provided that allow for nucleic acid amplification. In some aspects, the disclosure provides modified polymerases having lower systematic error as compared to a reference polymerase. In one aspect, the disclosure relates to modified polymerases useful for nucleic acid sequencing, genotyping, copy number variation analysis, paired-end sequencing and other forms of genetic analysis. In some aspects, the disclosure relates to modified polymerases useful for the generation of nucleic acid libraries or nucleic acid templates. In some aspects, the disclosure relates to the identification of homologous amino acid mutations that can be transferred across classes or families of polymerases to provide novel polymerases with altered properties.

Abstract: The present invention relates to variants of lysosomal acid lipase (LAL) and uses thereof.

Abstract: The present invention relates to polypeptides, nucleotides encoding the polypeptide, as well as methods of producing the polypeptides. The present invention also relates to detergent composition comprising polypeptides, a laundering method and the use of polypeptides.

Abstract: The present invention provides ISVD polypeptide derivatives capable of binding coagulation Factor IX(a) and Factor X(a) which are highly potent and provide a sufficiently long half-life such to allow for effective subcutaneous—as well as peroral administration. The ISVD polypeptides derivatives disclosed herein are thus suitable for treatment of haemophilia A, haemophilia A with inhibitors and acquired haemophilia A by various routes of administration including subcutaneous and peroral administration.

Abstract: The present disclosure provides, inter alia, a recombinant engineered deubiquitinase (DUB) and methods for treating or ameliorating an inherited ion channelopathy, such as long QT syndrome, Brugada syndrome, or cystic fibrosis, in a subject. Further provided are methods for screening mutations causing such inherited ion channelopathies for a trafficking-deficient mutation that is treatable by the recombinant engineered DUB disclosed herein.

Abstract: The present disclosure provides, inter alia, a recombinant engineered deubiquitinase (DUB) and methods for treating or ameliorating an inherited ion channelopathy, such as long QT syndrome, Brugada syndrome, or cystic fibrosis, in a subject. Further provided are methods for screening mutations causing such inherited ion channelopathies for a trafficking-deficient mutation that is treatable by the recombinant engineered DUB disclosed herein.

Abstract: Provided herein are polypeptides that are truncated modified recombinant forms of ADAMTS13, nucleic acid molecules encoding the polypeptides, drug delivery compositions and pharmaceutical compositions comprising the polypeptides, and methods of using the polypeptides and compositions (e.g., in the treatment of thrombotic thrombocytopenic purpura (TTP)).

Abstract: The present invention includes mutant AID, APOBEC, and Tet enzymes with improved functions. In one aspect the invention provides APOBEC fusion proteins comprising hyperactive deamination activity. In another aspect, the invention provides AID mutant proteins comprising hyperactive deamination activity. In yet another aspect, the invention provides mutant Tet proteins capable of stalling oxidation at a 5-hydroxymethylcytosine (hmC).

Abstract: Pathological retinal neovascularization is a common micro-vascular complication in several retinal diseases including retinopathy of prematurity (ROP), diabetic retinopathy, age related macular degeneration and central vein occlusion. Disclosed herein are compositions and methods useful for the treatment or prevention of retinal neovascularization and related diseases in a subject in need thereof. Exemplary methods include administering a composition including PEGylated arginase 1 to a subject in need thereof to promote reparative angiogenesis and decrease retinal neovascularization in the eye.

Abstract: The present invention relates to novel polypeptides capable of detoxifying Ochratoxin A (OTA) and methods (e.g., for detoxifying mycotoxins) based thereon. The present invention further relates to compositions, kits, transgenic plants, transgenic seeds, transgenic pollen grains, foodstuff, intermediate foodstuff; fodder, intermediate fodder; feed, intermediate feed; additive (e.g., foodstuff-, fodder- or feed additive), intermediate additive (e.g., foodstuff-, fodder- or feed intermediate additive); detoxifying agent, intermediate detoxifying agent; nutritional supplement, intermediate nutritional supplement, prebiotic, intermediate prebiotic and/or mixture/s thereof comprising one or more of the polypeptides capable of detoxifying Ochratoxin A (OTA).

Abstract: A number of T4 DNA ligase mutants exhibiting enhanced ligation activity in the presence of high salt concentrations compared to the wild-type ligase were engineered, characterized, and selected via gel electrophoresis of ligation products from a standard ligation assay. Ligase catalyzes the formation of phosphodiester bonds between the 5? and 3? ends of complementary cohesive ends or blunt ends of duplex DNA, a process that is vital to numerous molecular biology processes including cloning and sequencing.

Abstract: The present invention relates to chimeric (preferably, bifunctional) compounds, compositions comprising those compounds and methods of treating cancer in a patient or subject, especially including metastatic cancer where cancer cells exhibit overexpression (heightened expression) of cell surface urokinase-type plasminogen activator receptor (urokinase receptor) compared to normal (non-cancerous) cells. The compounds preferably covalently bind to the urokinase receptor and recruit native antibodies of the patient or subject where the antibodies can selectively degrade and/or deactivate targeted cancer cells through antibody-dependent cellular phagocytosis and/or antibody-dependent cellular cytotoxicity (ADCC) against a large number and variety of cancers, thus providing cancer cell death and/or an inhibition of growth, elaboration and/or metastasis of the cancer, including remission and cure of the patient's cancer.

Abstract: The present disclosure provides methods, compositions and systems for analyzing individual cells or cell populations through a partitioned analysis of contents of individual cells or cell populations, such as cancer cells and cells of the immune system. Individual cells or cell populations may be co-partitioned with processing reagents for accessing cellular contents, and for uniquely identifying the content of a given cell or cell population, and subsequently analyzing the content of the cell and characterizing it as having derived from an individual cell or cell population, including analysis and characterization of nucleic acid(s) from the cell through sequencing.

Abstract: The present invention relates to an RNA-mediated base editing system that enables recruitment of multiple effector proteins to a single target site, allowing for highly efficient base editing. The invention further relates to methods and kits for genetically modifying a cell.

Abstract: A method for high-throughput screening of a chimeric antigen receptor (CAR) cell library is provided comprising the steps of (a) providing a recognition sequence library, a hinge region sequence library, a transmembrane sequence library and an intracellular domain sequence library; (b) preparing a CAR library; (c) preparing a CAR-cell library by introduction to, and expression of, the plurality of CAR sequences of the CAR library in one or more cells or a cell line; (d) screening the CAR-cell library in an assay; (e) evaluating the at least one function of each member of the CAR-cell library; (f) obtaining one or more sequences of one or more CARs expressed in the CAR-cell library and linking the obtained sequence(s) to the at least one function of the members of the CAR-cell library; g) identifying and selecting the or each sequence based on function. Methods of preparing the CAR library and a CAR-cell library and uses thereof are also provided.

Abstract: The present invention relates to a molecular tracking device to follow distribution, acquisition, and retention of antigens in cells, such as in the lymphatic system of a subject. Molecular conjugates including an antigen conjugated to a nuclease-resistant tag are tracked using single-cell mRNA sequencing. The molecular tracking devices allow new approaches to investigate dynamic tissue dissemination of antigens and identify new mechanisms of antigen acquisition and retention at cellular resolution in vivo.

Abstract: Genetic circuits that control transgene expression in response to pre-defined transcriptional cues would enable the development of smart therapeutics. The present disclosure relates to engineered programmable single-transcript RNA sensors in which adenosine deaminases acting on RNA (ADARs) autocatalytically convert trigger hybridization into a translational output. This system amplifies the signal from editing by endogenous ADAR through a positive feedback loop. Amplification is mediated by the expression of a hyperactive, minimal ADAR variant and its recruitment to the edit site via an orthogonal RNA targeting mechanism. This topology confers high dynamic range, low background, minimal off-target effects, and a small genetic footprint. The circuits and systems disclosed herein leverage an ability to detect single nucleotide polymorphisms and modulate translation in response to endogenous transcript levels in mammalian cells.

Abstract: The present disclosure relates to a novel, engineered enveloped vector that can be used for gene delivery. The engineered enveloped vector comprises an engineered envelope comprising: (a) a viral envelope protein and optionally, (b) a non-viral membrane-bound protein. The present disclosure also provides a method of making and using the engineered enveloped vector.

Abstract: Methods of preparing a sequencing library includes fragmenting FFPE-extracted DNA into fragments about 800 bp in length on average; ligating adaptors to the fragments to form adaptor-ligated fragments; size-selecting the adaptor-ligated fragments to provide a mixture enriched for selected adaptor-ligated fragments with a size of about 600 to about 900 bp; and amplifying the selected adaptor-ligated fragments to obtain amplicons.

Abstract: The invention relates to barcoded nucleic acid nanostructure delivery compositions for in vivo screening for subsequent use in vivo therapeutic delivery, and methods therefor. More particularly, the invention relates to nucleic acid nanostructure delivery compositions, such as DNA origami structures, associated with barcodes for high throughput in vivo screening of the nucleic acid nanostructure delivery compositions for subsequent use in drug delivery, and methods therefor.

Abstract: This disclosure provides systems, methods, and compositions for site-specific genetic engineering using Programmable Addition via Site-Specific Targeting Elements (PASTE). PASTE comprises the addition of an integration site into a target genome followed by the insertion of one or more genes of interest or one or more nucleic acid sequences of interest at the site. PASTE combines gene editing technologies and integrase technologies to achieve unidirectional incorporation of genes in a genome for the treatment of diseases and diagnosis of disease.

Abstract: Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of ATXN1 RNA in a cell or subject, and in certain instances reducing the amount of ATXN1 in a cell or subject. Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a neurodegenerative disease. Such symptoms and hallmarks include gait and limb ataxia, cognitive impairments, difficulty with speaking and swallowing, atrophy of the cerebellum and brainstem in magnetic resonance imaging (MRI), neurochemical abnormalities in the cerebellum and brainstem detected via magnetic resonance spectroscopy (MRS), and death within 10-15 years of symptom onset. Such neurodegenerative diseases include Spinocerebellar ataxia type 1.

Abstract: Disclosed herein include microRNA antagonists, therapeutic compositions that include one or more of such microRNA antagonists, and methods of treating and/or ameliorating cardiac diseases and/or muscular dystrophy disorders with the microRNA antagonists. Also included are combination therapies, wherein a therapeutic composition disclosed herein and an additional therapy agent are provided to a subject having or suspected of having cardiac disease and/or muscular dystrophy disorder. In particular, some embodiments disclosed herein relate to compositions and methods for transiently administering a mixture of microRNA antagonists for promoting cardiomyocyte proliferation and cardiac regeneration.

Abstract: Disclosed herein are antisense compounds and methods for decreasing apo(a) to treat, prevent, or ameliorate diseases, disorders or conditions related to apo(a) or Lp(a). Certain diseases, disorders or conditions related to apo(a) or Lp(a) include inflammatory, cardiovascular and/or metabolic diseases, disorders or conditions. The antisense compounds disclosed herein can be used to treat such diseases, disorders or conditions in an individual in need thereof.

Abstract: The present disclosure provides pharmaceutic compositions for oral delivery comprising an antisense oligonucleotide (e.g., CIVI 008) and an oral delivery agent such as 5-CNAC. In some aspects, the disclosure provides a capsule comprising a dry blend of CIVI 008 and 5-CNAC, and optionally a statin.

Abstract: The present invention provides methods of treating or preventing chronic intestinal inflammation and/or inflammatory bowel disease (IBD) comprising administering GATA6-AS1 lncRNA to a patient in need thereof.

Abstract: This disclosure relates to novel targets for angiogenic disorders. Novel oligonucleotides are also provided. Methods of using the novel oligonucleotides for the treatment of angiogenic disorders (e.g., preeclampsia) are also provided.

Abstract: Provided herein are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas/guide RNA (gRNA) systems, for the transcriptional repression of Hepatitis B viral (HBV) genes to promote a cellular phenotype that leads to the reduction of HBV infection. In some embodiments, the epigenetic-modifying DNA-targeting systems bind to or target a target site of at least one gene or regulatory element thereof in a Hepatitis B viral DNA sequence in cell. In some aspects, the provided systems relate to the transcriptional repression of one or more Hepatitis B viral gene and/or regulatory element thereof. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in repressing Hepatitis B viral replication and expression in connection with Hepatitis B infections.

Abstract: Provided herein are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas/guide RNA (gRNA) systems, for the transcriptional repression of Hepatitis B viral (HBV) genes to promote a cellular phenotype that leads to the reduction of HBV infection. In some embodiments, the epigenetic-modifying DNA-targeting systems bind to or target a target site of at least one gene or regulatory element thereof in a Hepatitis B viral DNA sequence in cell. In some aspects, the provided systems relate to the transcriptional repression of one or more Hepatitis B viral gene and/or regulatory element thereof. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in repressing Hepatitis B viral replication and expression in connection with Hepatitis B infections.

Abstract: Compositions and methods are provided for the analysis and treatment of conditions relating to clonal hematopoiesis of indeterminate potential (CHIP). In some embodiments, treatment is provided to reduce the progression of CHIP, particularly to reduce the progression to hematologic malignancy and/or heart disease. In other embodiments, methods are provided for determining clonal expansion, for example as a molecular diagnostic test that enables determination of clonal growth rate from a single sample. The method for determining clonal expansion can be applied to identify factors that influence clonal expansion, including environmental, metabolic, microbiome, and genetic.

Abstract: The present disclosure relates to the technical field of genetic engineering, in particular to a target ligand. The target ligand provided by the present disclosure forms a siRNA conjugate with a specific small interfering RNA sequence, which targets to an angiopoietin like 3 (ANGPTL3), and reduce the expression quantity of an ANGPTL3 protein by degrading a transcript of an ANGPTL3 gene in a cell. Therefore, the siRNA conjugate formed by the target ligand provided by the present disclosure may be used to prevent and/or treat a dyslipidemia disease.

Abstract: The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the dipeptidyl peptidase 4 (DPP4) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a DPP4 gene and to methods of treating or preventing a DPP4-associated disease, such as metabolic diseases, e.g., diabetes or lipid metabolism diseases, in a subject.

Abstract: The present invention refers to oligonucleotides comprising 10 to 25 nucleotides, wherein at least one of the nucleotides is modified, and the oligonucleotide hybridizes with mRNA of angiotensin-converting enzyme 2 (ACE2) of SEQ ID NO.1 and/or with pre-mRNA of ACE2 of SEQ ID NO.2 resulting in a reduction of the level of ACE2, ACE2 mRNA, ACE2 pre-mRNA or a combination thereof of 30 to 99% compared to an untreated control. The invention is further directed to a pharmaceutical composition comprising such oligonucleotide. The oligonucleotide and the pharmaceutical composition are used in a method of preventing and/or treating a viral disease.