C12N9/02

NON-VIRAL DNA VECTORS AND USES THEREOF FOR EXPRESSING PHENYLALANINE HYDROXYLASE (PAH) THERAPEUTICS

The application describes ceDNA vectors having linear and continuous structure for delivery and expression of a transgene. ceDNA vectors comprise an expression cassette flanked by two ITR sequences, where the expression cassette encodes a transgene encoding PAH protein. Some ceDNA vectors further comprise cis-regulatory elements, including regulatory switches. Further provided herein are methods and cell lines for reliable gene expression of PAH protein in vitro, ex vivo and in vivo using the ceDNA vectors. Provided herein are method and compositions comprising ceDNA vectors useful for the expression of PAH protein in a cell, tissue or subject, and methods of treatment of diseases with said ceDNA vectors expressing PAH protein. Such PAH protein can be expressed for treating disease, e.g., Phenylketonuria (PKU).

Synthetic metabolic funneling for biochemical production

Certain embodiments provide a method for preparing a biochemical product (e.g., phenol, catechol, or muconic acid, or a salt thereof). For example, such methods include contacting a recombinant host having two or more recombinant pathways with a fermentable carbon source and growing the recombinant cell for a time sufficient to synthesize the product. In certain embodiments, each recombinant pathway: 1) is capable of producing the same final biochemical product; 2) comprises at least one gene encoding a polypeptide; 3) is derived from a different endogenous metabolite as its immediate precursor; and 4) converges to the same final product or the same intermediate metabolite.

ARTIFICIAL NON-CODING RNA MODULE FOR ENHANCING NITROGEN FIXATION ABILITY OF MICROORGANISMS

An artificial non-coding RNA (ncRNA) module constructed by a synthetic biology technique and the use of the artificial ncRNA module in the construction of an artificial nitrogen fixation system are disclosed. The RNA module can enhance the post-transcriptional stability of nifHDK mRNA by interacting with a nitrogenase coding gene nifHDK mRNA, thereby improving the nitrogen fixation ability of a chassis microorganism. A fusion expression vector carrying the artificial RNA module is constructed and transformed into different chassis nitrogen-fixing microorganisms. It is confirmed through experiments that, under nitrogen fixation conditions, the artificial RNA module of the present disclosure can significantly improve the nitrogenase activity of a recombinant engineering bacterial strain.

METHODS AND COMPOSITIONS FOR THE PRODUCTION OF ACETYL-COA DERIVED PRODUCTS
20230227864 · 2023-07-20 · ·

The present disclosure is related to genetically engineered microbial strains and related bioprocesses for the production of products from acetyl-CoA. Specifically, the use of dynamically controlled synthetic metabolic valves to reduce the activity of certain enzymes, leads to increased product production in a two-stage process.

Wheat with reduced lipoxygenase activity

A series of independent human-induced non-transgenic mutations found at one or more of the Lpx genes of wheat; wheat plants having these mutations in one or more of their Lpx genes; and a method of creating and finding similar and/or additional mutations of Lpx by screening pooled and/or individual wheat plants. The wheat plants disclosed herein exhibit decreased lipoxygenase activity without having the inclusion of foreign nucleic acids in their genomes. Additionally, products produced from the wheat plants disclosed herein display increased oxidative stability and increased shelf life without having the inclusion of foreign nucleic acids in their genomes.

BIOCATALYSTS AND METHODS FOR HYDROXYLATION OF CHEMICAL COMPOUNDS

The present disclosure provides engineered proline hydroxylase polypeptides for the production of hydroxylated compounds, polynucleotides encoding the engineered proline hydroxylases, host cells capable of expressing the engineered proline hydroxylases, and methods of using the engineered proline hydroxylases to prepare compounds useful in the production of active pharmaceutical agents.

ORGANELLE-TARGETED GENETICALLY-ENCODED VOLTAGE INDICATORS AND METHODS OF USE THEREOF
20230228737 · 2023-07-20 ·

As described herein, a hybrid voltage sensor genetically-encoded voltage indicator (GEVI) for mitochondria or endoplasmic reticulum includes a transmembrane domain, and a fluorescent protein, wherein a terminus of the transmembrane domain and a terminus of the fluorescent protein are covalently linked directly or by a linker comprising 1 to 20 amino acids, and wherein the transmembrane domain comprises SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4 or a peptide with greater than 85%, 90%, 95% or 98% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. Also described are expression vectors, expression cassettes, and organelle membranes, as well as methods of determining the voltage across an organelle using the GEVIs.

STEVIOL GLYCOSIDE TRANSPORT

The disclosure provides a recombinant cell capable of producing a steviol glycoside, wherein the cell comprises a nucleic acid coding for a variant of a parent polypeptide, wherein the variant has steviol glycoside transport mediating activity, wherein the variant comprises an amino acid sequence which, when aligned with the amino acid sequence of the parent polypeptide, comprises at least one modification of the amino acid residue corresponding to any of the amino acids in the amino acid sequence of the parent polypeptide, wherein the variant has an improved ability to produce rebaudioside M and optionally other steviol glycosides extracellularly if compared with the parent polytpeptide when measured under the same conditions.

Modified Microorganisms and Production Method of Compounds

A modified microorganism containing a genetic modification that suppresses a transcription factor that controls expression of pyruvate dehydrogenase, in which the microorganism has and a production pathway of a C6 compound, and in which the C6 compound is at least one compound selected from the group consisting of adipic acid, hexamethylenediamine, 1,6-hexanediol, 6-aminohexanoic acid, 6-amino-1-hexanol, 6-hydroxyhexanoic acid, 3-oxoadipic acid, 3-hydroxyadipic acid, and 2,3-dehydroadipic acid.