Abstract: The present disclosure provides an automated method of producing genetically modified immune cells, including chimeric antigen receptor T (CAR T) cells, utilizing a fully-enclosed cell engineering system.

Abstract: Methods and cell compositions are provided in which expression of BHLHE40 is modulated to direct T cell phenotype and function.

Abstract: Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), for example.

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: A composition is for the targeted knockout of a gene on double-stranded DNA in a biological cell. A method is for the targeted knockout of a gene on double-stranded DNA in a biological cell. A preparation includes a biological cell prepared in vitro. The biological cell includes a gene on double-stranded DNA, which is knocked-out in a targeted manner. A kit is for the targeted knockout of a gene on double-stranded DNA in a biological cell. Another method is for treating a subject afflicted with a disease associated with a mutated gene. Nucleic acid molecules can be a component of the composition and methods.

Abstract: The present invention relates to a preparation method for a self-organized cardiac organoid. The invention also relates to a cardiac tissue organoid obtained by the method and to its use in regenerative medicine, as a tissue implant, or in drug screening.

Abstract: Human skeletal muscle stem cells were generated from facioscapulohumeral muscular dystrophy (FSHD) and healthy control iPSC using a transgene-free skeletal muscle differentiation protocol and production of stable iMyoblasts. Analyses revealed that FSHD and healthy control iMyoblasts are embryonic-like myogenic cells that undergo myotube differentiation ex vivo by growth factor depletion and are efficiently transplantable into the tibialis anterior (TA) muscles of NSG mice, where human muscle under-goes embryonic-to-adult myosin isoform switching. The DUX4 FSHD disease gene maintains its hypomethylated disease state inFSHD iPSC and iMyoblast, and its expression is upregulated during myotube differentiation and in muscle xenografts. Consequently, these iMyoblasts accurately exhibit the molecular pathology of human muscular dystrophies and are useful for the development of drug, gene editing and stem cell therapeutics.

Abstract: Provided are methods and systems for an enriched production of high-quality extracellular vesicles (EVs) from a mammalian cell. In some cases, the methods may comprise culturing the cell in a chemically-defined protein-free (CDPF) medium with the addition of a lysosome inhibitor to increase production of EVs. In some cases, the CDPF medium is supplemented with additives.

Abstract: Provided are a recombinant CHO cell, a construction method therefor, a detection system containing same, and a detection method using same. The recombinant CHO cell stably expresses NGF on the cell surface thereof and can be used as a target cell for an ADCC effect assay and a CDC effect assay.

Abstract: The present invention provides compositions comprising high densities of endothelial cells (such as human umbilical vein endothelial cells) in freezing media, methods of producing such compositions, and methods of using such compositions in the preparation of therapeutic endothelial cell compositions. Such compositions and methods provide numerous advantages including eliminating the requirement to remove cryopreservatives before administration of therapeutic endothelial cell compositions to human subjects and requiring minimal manipulations and human interventions before use in therapeutic methods.

Abstract: There is provided a device for vascularising a cell aggregate, the device comprising: a matrix region configured to contain a gel-like matrix and cells that form a vasculature network within th ematrix region. The matrix region having at least one opening for positioning the cell aggregate therein based on a desired three-dimensional spatial location; and one or more fluidic regions configured to contain a supporting fluid that is capable of supporting vascularisation of the cell aggregate, the one or more fluidic regions being in fluid communication with the matrix region, wherein a flow passage from the one or more fluidic regions to a gel-like matrix disposed in the matrix region is configured to allow three-dimensional vascularisation around the cell aggregate and perfusion of the vasculature once formed. There is also provided a chip comprising a plurality of the device as disclosed herein and a method for vascularising a cell aggregate.

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: Disclosed are findings that: (a) induced pluripotent stem cells derived from aged donors (A-iPSC) show increased genomic instability, a defect in apoptosis, a defect in glucose metabolism, and a blunted DNA damage response are compared to those derived from young donors (Y-iPSC); and (b) inhibition of excessive glutathione-mediated H202 scavenging activity, found to be associated with A-iPSC and in turn inhibiting DNA damage response and apoptosis, substantially rescues these defects and reduces the oncogenic potential of A-iPSC. Supplementation of pluripotency factor ZSCAN 10 (shown to be poorly activated in A-iPSC and to act upstream of glutathione involvement), e.g.

Abstract: The present invention provides compositions and methods for reprogramming somatic cells using purified RNA preparations comprising single-strand mRNA encoding an iPS cell induction factor. The purified RNA preparations are preferably substantially free of RNA contaminant molecules that: i) would activate an immune response in the somatic cells, ii) would decrease expression of the single-stranded mRNA in the somatic cells, and/or iii) active RNA sensors in the somatic cells. In certain embodiments, the purified RNA preparations are substantially free of partial mRNAs, double-stranded RNAs, un-capped RNA molecules, and/or single-stranded run-on mRNAs.

Abstract: The disclosure is directed to a method for seed train expansion of adherent cells comprising culturing cells with a serum-supplemented growth medium in a N-2 vessel; removing the cells from the serum-supplemented medium; inoculating the cells from step into a serum-free growth medium in a N-1 vessel; culturing the cells in the N-1 vessel under suspension conditions; and inoculating a growth medium in a bioreactor with the suspension-cultured cells. In some aspects, the adherent cells are not suspension-adapted. In some aspects, the adherent cells are suspension-adapted. In some aspects, the adherent cells produced by the seed train expansion method are used to produce viral vectors. In some aspects, the viral vectors are AAV vectors.

Abstract: The present invention relates to an oncolytic virus comprising: (i) a fusogenic protein-encoding gene and (ii) an immune stimulatory molecule-encoding gene.

Abstract: The present disclosure relates to engineered ketoreductase polypeptides for the preparation of hydroxyl substituted carbamate compounds, and polynucleotides, vectors, host cells, and methods of making and using the ketoreductase polypeptides.

Abstract: Mutant polymerases are provided that have improved ability to incorporate modified nucleotides, including 3?-OH unblocked reversible terminators. The mutant polymerases may be used in a variety of applications, such as for polynucleotide sequencing, primer extension reactions, and template-independent enzymatic oligonucleotide synthesis.

Abstract: Disclosed are methods for purifying immobilized lipase preparations from various pigments and colorants as well as other contaminants, using various lipase-catalyzed reactions with oils and fats as substrates, such as esterification, transesterification and interesterification reactions. Also disclosed are purified immobilized lipase preparations and their industrial uses.

Abstract: The present invention relates to: a prime editing composition comprising a prime editor protein or a nucleic acid encoding same, and a prime editing guide RNA (pegRNA); and a prime editing method.

Abstract: The present disclosure provides compositions, systems, and methods for genome editing, efficient knock-in of large DNA fragments, and long-term, stable, high expression of integrated transgenes. Also provided are modified cells, vaccines comprising modified cells, and methods of using such cells to induce an immune response.

Abstract: The invention provides for systems, methods, and compositions for targeting nucleic acids. In particular, the invention provides non-naturally occurring or engineered RNA-targeting systems comprising a novel RNA-targeting Cas13b effector protein and at least one targeting nucleic acid component like a guide RNA or crRNA.

Abstract: The present disclosure encompasses engineered meganucleases that bind and cleave recognition sequences within a dystrophin gene. The present disclosure also encompasses methods of using such engineered meganucleases to make genetically modified cells. Further, the disclosure encompasses pharmaceutical compositions comprising engineered meganuclease proteins, or polynucleotides encoding engineered meganucleases of the disclosure, and the use of such compositions for the modification of a dystrophin gene in a subject, or for treatment of Duchenne Muscular Dystrophy.

Abstract: Compositions and methods for editing, e.g., introducing double-stranded breaks, within the KLKB1 gene are provided. Compositions and methods for treating subjects having hereditary angioedema (HAE), are provided.

Abstract: Compositions and methods are provided for variant Cas systems and elements comprising such systems, including, but not limiting to, Cas endonuclease variants, guide polynucleotide/Cas endonuclease complexes comprising Cas endonuclease variants, as well as guide polynucleotides and guide RNA elements that can interact with Cas endonuclease variants. Compositions and methods are provided for genome modification of a target sequence in the genome of a cell. The methods and compositions employ a guide polynucleotide/Cas endonuclease system comprising a Cas9 endonuclease variant to provide an effective system for modifying or altering target sequences within the genome of a cell or organism.

Abstract: The present disclosure provides codon optimized nucleotide sequences encoding hum n alpha-galactosidase A, vectors, and host cells comprising codon optimized alpha-galactosidase A sequences, and methods of treating disorders such as Fabry disease comprising administering to the subject a codon optimized sequence encoding human alpha-galactosidase A.

Abstract: The present invention relates to fusion proteins of ACE2 with IgG Fc and the medical use of these fusion proteins, in particular in the prevention or treatment of infections with coronaviruses such as SARS-CoV-2.

Abstract: The specification discloses Clostridial toxins or Clostridial toxin chimeras comprising an inactivation cleavage site, polynucleotide molecules encoding such toxins or chimeras, compositions comprising such toxins or chimeras, and method of producing such toxins or chimeras.

Abstract: An isolated nucleic acid that includes an open reading frame encoding a lanthipeptide protease polypeptide for scarless tag removal from a polypeptide is presented. Reagents, expression constructs and methods are also provided for preparing a scarless tag polypeptide product from a tagged polypeptide precursor containing a lanthipeptide protease cleavage site. The reagents are directed to novel lanthipeptide proteases and expression constructs and polypeptide precursors that include highly specific lanthipeptide protease substrate recognition sequence. Methods are provided that enable scarless tag removal from a cognate lanthipeptide, a non-cognate lanthipeptide or a heterologous polypeptide that includes extraneous amino acid sequences, such as leader peptides and tags.

Abstract: Provided are MTSP-1 polypeptides modified to have altered activity and/or specificity so that they cleave a complement protein, such as complement protein C3, to inhibit its activity and thereby inhibit complement activation. The modified MTSP-1 polypeptides that inhibit complement activation can be used for treatment of diseases and conditions in which complement activation plays a role. Such diseases and conditions include inflammatory diseases and diseases with an inflammatory component. Exemplary of these disorders are ischemic and reperfusion disorders, including myocardial infarction and stroke, sepsis, autoimmune diseases, ophthalmic disorders, such as diabetic retinopathies and macular degeneration, including age-related macular degeneration (AMD), and transplanted organ rejection, such as renal delayed graft function (DGF).

Abstract: The present invention provides a method for improving or controlling the plasma half-life and/or bio-availability of blood coagulation factor IX (FIX), the method comprising modifying the GLA domain. Examples of such modifications include: (i) non-covalent bonding of a GLA-domain-recognizing antibody or an antibody fragment thereof to the GLA domain; (ii) reduced number of Gla residues in the GLA domain, in comparison to that of a native FIX; (iii) either or both of deletion of one or more glutamic acid residues in the GLA domain and substitution of one or more glutamic acid residues in the GLA domain with another amino acid; and (iv) deletion of a part or all of the GLA domain. The present invention also provides a FIX with improved pharmacokinetics which carries such modifications, a pharmaceutical composition containing the FIX as an active ingredient, a method for producing the FIX, and such.

Abstract: The present invention provides engineered leucine decarboxylase (LDC) polypeptides and compositions thereof, as well as polynucleotides encoding the engineered leucine decarboxylase (LDC) polypeptides. In some embodiments, the engineered LDC polypeptides are optimized to provide enhanced catalytic activity, as well as reduced sensitivity to proteolysis, and/or increased tolerance to low pH environments. In some embodiments, the engineered LDC polypeptides are optimized to provide improved storage stability. The present invention also provides methods for the use of the compositions comprising the engineered LDC polypeptides for therapeutic and industrial purposes.

Abstract: Enzyme mutant with squalene-hopene-cyclase activity, selected from mutants of a wild-type enzyme comprising an amino acid sequence selected from SEQ-ID No: 1 to 3 or an amino acid sequence derived therefrom with a degree of sequence identity in the range of from 60 to 99.9% of SEQ-ID No. 1 to 3, wherein the mutant catalyzes a one-step monocyclization reaction to produce products such as gamma-dihydroionone and/or alpha-dihydroionone.

Abstract: Apparatuses and methods are described herein for processing polynucleotides in a sealed path environment. The apparatuses include optical sensors to monitor operations and to track material usage for good manufacturing practice.

Abstract: Apparatuses and methods are described herein for processing polynucleotides in a sealed path environment. The apparatuses include optical sensors to monitor operations and to track material usage for good manufacturing practice.

Abstract: The present invention relates to a method for determining an extent of cyclisation of a peptide ligand displayed on a genetic display system, wherein the peptide ligand comprises a polypeptide covalently linked to a molecular scaffold at two or more amino acid residues, comprising the steps of exposing the polypeptide displayed on the genetic display system to the molecular scaffold, wherein said polypeptide comprises two or more peptide reactive groups on said two or more amino acid residues which form covalent bonds with the molecular scaffold at two or more scaffold reactive groups, to give the peptide ligand; removing unreacted molecular scaffold from the genetic display system; exposing the peptide ligand displayed on the genetic display system to a first probe, wherein the first probe binds to a first unconjugated reactive group on the peptide ligand; and measuring the first unconjugated reactive group on the peptide ligand.

Abstract: The present invention provides methods and kits for identifying interactions between a test compound and a phage-displayed polypeptide, wherein the phage-displayed polypeptide is not a kinase but a protein domain of interest, such as an SH2 domain of interest. The protein domain of interest is displayed on the phage, and the interactions are evaluated in the presence of a reference ligand and in the presence and absence of the test compound.

Abstract: The present invention relates to preparation, sequencing and analysis of a sequencing library of adaptor-tagged fragments, wherein the fragments have different orientations relative to a sequencing adaptor.

Abstract: Array-based enzymatic oligonucleotide synthesis creates a large number of polynucleotides using an uncontrolled and template independent polymerase such as terminal deoxynucleotidyl transferase (TdT). Spatial control of reaction conditions on the surface of the array allows creation of polynucleotides with a variety of arbitrary sequences. Spatial control may be implemented by removing protecting groups attached to nucleotides only at a selected location on the array or by other techniques such as location-specific regulation of enzymatic activity. The ratio of polynucleotides with protecting groups to unprotected polynucleotides used during a cycle of synthesis is adjusted to control the length of homopolymers created by the polymerase. Digital information may be encoded in the enzymatically synthesized polynucleotides.

Abstract: A method for analyzing cell epigenomics from multiple dimensions. The method comprises the following steps: by using ChiTag transposase and conventional Tn5 transposase in cells, respectively embedding different linker sequences, and achieving common analysis of information of a chromatin open region and information of a specific protein binding sequence on a cellular level. The method has important application prospects in aspects such as study of development and/or disease related cell population heterogeneity, drawing of a cell map, analysis of tumor cells having different clinical characteristics, and clinical study of evolution and/or metastasis of tumor cells.

Abstract: The disclosure provides artificial nucleic acid introns configured for selective splicing in cells with aberrant RNA splicing activity, e.g., neoplastic cells. The artificial intron can comprise a 5? splice site, a canonical 3? splice site, at least one cryptic 3? splice site, a pyrimidine-rich domain, and at least one branchpoint. Also provided are constructs integrating the artificial introns with exons in a configuration that, when the artificial intron is spliced out by the aberrant RNA splicing factors, encode a functional protein. Also disclosed are methods that employ the disclosed platform of selective expression, including, targeted gene therapy methods (e.g., in cancers), diagnostics and imaging, and drug screening.

Abstract: The present invention provides methods for large-scale production of a composition enriched for full-length mRNA molecules using an SP6 RNA polymerase and compositions produced using such methods and uses thereof.

Abstract: The present invention relates to RNAi agents, e.g., dsRNA agents, targeting the complement factor B (CFB) gene. The invention also relates to methods of using such RNAi agents to inhibit expression of a CFB gene and to methods of treating or preventing a CFB-associated disease in a subject.

Abstract: A single-stranded antisense oligonucleotide, or a pharmaceutically acceptable salt thereof, capable of modulating expression and/or function of RPS25 gene, wherein nucleotides of the single-stranded antisense oligonucleotide are bonded to each other via a phosphate group and/or a modified phosphate group, the single-stranded antisense oligonucleotide includes a gap region, a 3? wing region bonded to a 3? end of the gap region, and a 5? wing region bonded to a 5? end of the gap region, the gap region is a deoxyribose-based nucleic acid optionally including a nucleic acid having a modified sugar moiety, each of the 3? wing region and the 5? wing region is a modified nucleotide, the single-stranded antisense oligonucleotide has a base length of 12- to 30-mer, and a base sequence of the antisense oligonucleotide is: a base sequence with a sequence identity of 90% to 100% to a base sequence complementary to at least one target region of the same base length as the antisense oligonucleotide present in the base sequ

Abstract: The invention features a method of treating a disease or disorder in a subject, the method comprising administering a therapeutically effective amount of a 5?-tiRNA to treat the disease or disorder in the subject.

Abstract: The present disclosure relates to compositions of matter and assay methods used to detect one or more target nucleic acids of interest in a sample. The compositions and methods allow one to control reaction kinetics of the cascade assay by two orders of magnitude via molecular design of one of the reaction components; further, varying molecular design also allows for quantification of target nucleic acids of interest over a large range of concentrations or discriminating between extremely low copy numbers of target nucleic acids of interest.

Abstract: The invention relates to modified oligonucleotides, e.g., saRNAs useful in upregulating the expression of a target gene and therapeutic compositions comprising such oligonucleotides. Methods of using the oligonucleotides and the therapeutic compositions are also provided.

Abstract: Synthetic sRNA for inhibiting prokaryote gene expression is described, which includes (i) an Hfq binding site and (ii) a region that forms a complementary bond with a target gene mRNA. Vectors encoding the synthetic sRNA are described, as well as recombinant prokaryotes transformed with such vectors, methods of inhibiting prokaryote gene expression, and methods of gene screening and strain improvement. The synthetic sRNA is able to control single and multiple target genes at a time, and is particularly useful for inhibiting a gene expression of Gram-positive bacteria, e.g., in a recombinant Corynebacterium for mass production of high value products that do not require fossil fuels with associated environmental problems.

Abstract: SnRNA systems comprising engineered stem loops are disclosed herein.

Abstract: Described herein are oligonucleotides (e.g., single-stranded oligonucleotides) and compositions thereof for targeting a mutation in the spliceosome, such as the U 1 small nuclear RNA (snRNA), as well as related methods of use.