Disclosed herein are methods for N-demethylating an N-methylated compound using an enzymatic reaction, rather than, e.g. a chemical modification. Also provided herein are enzymes for performing the reaction.

The present disclosure provides recombinant nucleic acids comprising one or more polynucleotides encoding a Serine Protease Inhibitor Kazal-type (SPINK) polypeptide (e.g., a SPINK5 polypeptide); viruses comprising the recombinant nucleic acids; compositions and formulations comprising the recombinant nucleic acids and/or viruses; methods of their use (e.g., for the treatment of Netherton Syndrome); and articles of manufacture or kits thereof.

The present invention relates to a genetically modified human stem cell, in which said human stem cell comprises an exogenous nucleic acid comprising a region coding for a fusion protein comprising a mutant human cytochrome P450 2B6 (CYP2B6*) protein of SEQ ID No. 1 or a variant or fragment thereof and an NADPH-cytochrome P450 reductase protein of SEQ ID No. 2 or a variant or fragment thereof, functionally bound to a promoter, said exogenous nucleic acid having been integrated into one of the genomic safe harbors of said human stem cell. The invention also relates to the use of said cell for the treatment of cancer and/or cancer recurrences and/or associated metastases, particularly the solid tumours, and in particular the hepatocellular carcinomas and/or associated metastases.

Described herein are production cells, and methods for identifying, selecting, or culturing production cells comprising tyrosine auxotrophy selection marker system, based on a combination of sequence encoding a phenylalanine hydroxylase (PAH) which lacks a functional N-terminal regulatory domain, and a sequence encoding a GTP cyclohydrolase 1 (GCH1). Also described are methods of making a production cell and making a product with said production cell.

Glucuronosyltransferase 1 gene which catalyzes glucuronic acid transfer to the hydroxyl group at the 3-position in an oleanane-type triterpenoid is identified. Glucuronosyltransferase 1 gene having a desired activity, derived from a Fabaceae plant (soybean, Glycyrrhiza, and Lotus japonicus), and containing nucleotide sequences represented by SEQ ID Nos: 2, 4, and 6, respectively, is provided.

The invention relates to compositions and methods for making and use of a real-time cellular sensor. Components of a multipart enzyme are sequestered in different cellular compartments and only come together after receptor activation; a pool of substrate is made available in the cell to ensure real-time enzymatic output.

Methods for recombinant production of steviol glycoside and compositions containing steviol glycosides are provided by this invention.

A gene editing system is provided that comprises a first nucleotide molecule encoding a dCas9-Ten-Eleven Translocation methylcytosine dioxygenase 1 catalytic domain (TET1CD) fusion protein; and a second nucleotide molecule encoding at least one small guide RNA (sgRNA), comprising: a scaffold region and a spacer region, wherein the spacer region hybridizes to a nucleotide sequence complementary to a target sequence adjacent to a 5′-end of a protospacer adjacent motif (PAM), and wherein the target sequence and the PAM are located within 1 kilobase (kb) of the transcriptional start site (TSS) of the CDKL5 gene. Methods of making and using the system are further described herein.

A method for re-expression of hypermethylated RASAL1, hypermethylated LRFN2, and hypermethylated KLOTHO based on an inactivated CRISPR-based system and a DNA dioxygenase as well as a gRNA guiding the construct to the RASAL1, LRFN2, and KLOTHO gene for demethylation of hypermethylated RASAL1, hypermethylated LRFN2, and hypermethylated KLOTHO, in particular, hypermethylated RASAL1, LRFN2, and KLOTHO promoter, thus, allowing re-expression of RASAL1, LRFN2, and KLOTHO for the treatment of fibrosis, cancer or neuronal disorders in a subject is provided. A kit of parts for allowing re-expression of hypermethylated RASAL1, hypermethylated LRFN2, and hypermethylated KLOTHO in a subject, a vector or vector system, and nucleic acid constructs are also provided.

The present invention is drawn to artificial metalloenzymes for use in cyclopropanation reactions, amination and C—H insertion.