Abstract: Disclosed herein is a method for producing a sialylated N-glycosylated recombinant protein in periplasm of a recombinant Escherichia coli. The sialylated N-glycosylated recombinant protein is produced in the periplasm of a recombinant Escherichia coli strain W3110?nanKETA::Kan, and a sialylated oligosaccharide chain is Neu5Ac-?-2,6-Gal-?-1,4-GlcNAc-?-1,3-Gal-?-1,3-GlcNAc.

Abstract: Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of PRNP RNA in a cell or animal, and in certain instances reducing the amount of PrP protein in a cell or animal. Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a neurodegenerative disease. Such symptoms and hallmarks spongiform changes in the brain, development of abnormal protein aggregates, neuronal loss, markers of neuronal loss, rapidly progressing dementia, and death. Such neurodegenerative diseases include prion diseases, Creutzfeldt-Jakob disease (CJD), variant Creutzfeldt-Jakob Disease (vCJD), familial Creutzfeldt-Jakob Disease (fCJD), Gerstmann-Straussler-Scheinker syndrome, fatal familial insomnia, kuru, Alzheimer's disease, or Parkinson's disease.

Abstract: In various aspects and embodiments the invention provides a method of treating epilepsy in a subject in need thereof, the method comprising contacting a target cell of the subject with an effective amount of an HCN4 disrupting agent.

Abstract: Provided are methods, compositions, recombinant DNA molecules, and kits for cloning T cell receptors (TCRs). The methods facilitate construction of TCR expression libraries from biological samples containing antigen-specific T cells, including but not limited to tumor biopsies, including frozen tumor biopsies. Peripheral T cells that were engineered with library-derived TCR genes show potent therapeutic anti-tumor effects. The method can be performed using any sample that contains T cells, and can be performed with oligoclonal populations of T cells, such as T cells that have infiltrated a tumor. Primer combinations for first strand cDNA synthesis, second strand cDNA synthesis, and for cloning a plurality of distinct TCR ? and TCR ? chains into a plurality of vectors are provided. Cells containing the vectors are provided, as are kits for use in rapid cloning of the TCR ? and TCR ? chains.

Abstract: The present disclosure provides variant type V CRISPR/Cas effector polypeptides, fusion polypeptides comprising the variant type V CRISPR/Cas effector polypeptides, and nucleic acids comprising nucleotide sequences encoding the variant polypeptides and fusion polypeptides. The present disclosure provides methods of binding, or binding and nicking, a target nucleic acid, using a variant type V CRISPR/Cas effector polypeptide of the present disclosure. The present disclosure provides methods of detecting a single-stranded DNA, using a variant type V CRISPR/Cas effector polypeptide of the present disclosure.

Abstract: Proposed is a genomic editing vector for Eubacterium callanderi for the CRISPR/Cas9 system. The genomic editing vector for Eubacterium callanderi includes a DNA sequence encoding a guide RNA (gRNA) of a cleavage target gene; a DNA sequence encoding a Cas9 protein; a Prbo promoter that is operably linked to the DNA sequence encoding a Cas9 protein; a replication starting point derived from E. coli; a replication starting point for Eubacterium callanderi; and a marker for selecting transgenic strains. It is possible to provide a genomic editing vector for Eubacterium callanderi that can be applied to Eubacterium callanderi strains that are acetogen that is suitable for syngas biorefinery, a method for editing a genome of Eubacterium callanderi strains using the same, and transgenic Eubacterium callanderi strains using the same.

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 is a genome editing composition for prime editing including a prime editor protein and a prime editing guide RNA, for editing a Z-type mutation of alpha-1 antitrypsin deficiency. The composition according to an aspect includes both a pegRNA sequence capable of effectively editing the SERPINA1 gene and prime editor 2 (PE 2), and thus, may effectively deliver the prime editor to a cell, and act specifically for a target sequence of the SERPINA1 gene, enabling genome editing with high accuracy, and therefore, may be useful as a SERPINA1 gene editing platform. In addition, the composition is capable of correcting the Z-type mutation of the SERPINA1 gene, and may be used for treatment or prevention of alpha-1 antitrypsin deficiency.

Abstract: Disclosed herein are compositions of transcription activator-like effectors transcription factors and methods of using the compositions for inducing gene expression of mammalian genes.

Abstract: The invention pertains to an inducible CRISPR system for controlling expression of a CRISPR complex with an inducible fusion promoter. One embodiment of the invention provides HIV LTR-minimal Drosophila hsp70 fusion promoter that can be used for inducible co-expression of gRNA and Cas9 in HIV-infected cells to target cellular cofactors such as Cyclin T1. A single introduction of such embodiment leads to sustained suppression of HIV replication in stringent, chronically infected HeLa-CD4 cell lines as well as in T-cell lines. In another embodiment, the invention further relates to enhancement of HIV suppression by incorporating cis-acting ribozymes immediately upstream of the gRNA in the inducible CRISPR system construct. The inducible fusion promoter is adaptable for other tissue- or cell-type specific expression of the inducible CRISPR system.

Abstract: Reversibly blocked nucleoside analogues and methods of using such nucleoside analogues for sequencing of nucleic acids are provided.

Abstract: Presented herein are methods and compositions for tagmentation of nucleic acids. The methods are useful for generating tagged DNA fragments that are qualitatively and quantitatively representative of the target nucleic acids in the sample from which they are generated.

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

Abstract: Novel intron fragments are provided. The intron fragments can increase gene expression to levels equal to or higher than those achieved by the full-length intron while maintaining their ability to increase gene expression even when combined with various types of promoters and splicing donors. Particularly, the intron fragments enable loading of larger transgenes when used in genetic information delivery systems whose size is limited, for example, adeno-associated viruses (AAVs) and rhabdoviruses. Therefore, the use of the intron fragments is expected to extend the range of therapeutic genes.

Abstract: Fusion proteins comprising a DNA binding domain, e.g., a TAL effector repeat array or zinc finger, and a catalytic domain comprising a sequence that catalyzes hydroxylation of methylated cytosines in DNA, and methods of use thereof.

Abstract: The present invention relates to a method of editing a filamentous fungal genome by direct introduction of a genome editing protein molecule or complex. The method includes three modes. In the first mode, a genome editing protein molecule or complex for a target gene is directly introduced into a cell of an Aspergillus fungus, to edit a gene in the Aspergillus fungal genome. In the second mode, a genome editing protein molecule or complex for a target region in a filamentous fungal genome, and a desired DNA fragment, are directly introduced into a filamentous fungal cell, to knock-in the DNA fragment to a desired target site in the filamentous fungal genome. In the third mode, genome editing protein molecules or complexes for plural target genes are directly introduced into a filamentous fungal cell, to carry out simultaneous editing of the plural genes in the filamentous fungal genome.

Abstract: Provided are methods, compositions, reagents, kits that are useful for detecting a specific nucleic acid region in genome with high efficiency and high sensitivity.

Abstract: Unbiased, genomewide and highly sensitive methods for detecting mutations, e.g., off-target mutations, induced by engineered nucleases.

Abstract: Compositions and methods for using programmable DNA binding proteins to increase the efficiency and/or specificity of targeted genome modification or to facilitate the detection of specific genomic loci in eukaryotic cells.

Abstract: Compositions and methods for using nucleosome interacting protein domains to increase accessibility of programmable DNA modification proteins to target chromosomal sequences, thereby increasing efficiency of targeted genome/epigenetic modification in eukaryotic cells.