The invention relates to mRNA therapy for the treatment of Acute Intermittent Porphyria (AIP). mRNAs for use in the invention, when administered in vivo, encode human porphobilinogen deaminase (PBGD), isoforms thereof, functional fragments thereof, and fusion proteins comprising PBGD. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to affect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of PBGD expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic metabolites associated with deficient PBGD activity in subjects, namely porphobilinogen and aminolevulinate (PBG and ALA).

The present invention relates to a microorganism variant having the ability to extracellularly produce heme, and more particularly to a metabolically engineered microorganism variant having the ability to extracellularly produce heme and a method of producing heme using the same. According to the present invention, heme, an organometallic compound which is increasingly used as a health food or food supplement for the treatment of porphyria, can be extracellularly secreted and produced in high yield using the microorganism variant, but not conventional chemical synthesis or enzymatic synthesis.

The invention relates to a genetically modified prokaryotic cell capable of improved iron-sulfur cluster delivery, characterized by a modified gene encoding a mutant Iron Sulfur Cluster Regulator (IscR) and one or more transgenes or upregulated endogenous genes encoding iron-sulfur (Fe—S) cluster polypeptides or proteins that catalyze complex radical-mediated molecular rearrangements, electron transfer, radical or non-redox reactions, sulfur donation or perform regulatory functions. The prokaryotic cells are characterized by enhanced activity of these iron-sulfur (Fe—S) cluster polypeptides, enhancing their respective functional capacity, and facilitating enhanced yields of compounds in free and protein-bound forms, including heme, hemoproteins, tetrapyrroles, B vitamins, amino acids, δ-aminolevulinic acid, biofuels, isoprenoids, pyrroloquinoline quinone, ammonia, indigo, or their precursors, whose biosynthesis depends on their activity. The invention further relates to a method for producing said compounds or their precursors using the genetically modified prokaryotic cell of the invention, and the use of the genetically modified prokaryotic cell.

The present disclosure provides compositions comprising a gene-editing polypeptide, a single-stranded donor DNA, and one or more staple oligonucleotides. The present disclosure provides compositions comprising a DNA nanostructure and a gene-editing polypeptide. The present disclosure provides gene editing methods using the compositions. The present disclosure provides methods of using the compositions to produce a genetically modified cell. The present disclosure provides kits useful for carrying out gene editing.

The present disclosure relates to human porphobilinogen deaminase derived proteins and polynucleotides and methods of using these proteins and polynucleotides.

The present invention relates to compositions of small activating nucleic acid molecules for increasing the expression of HMBS gene and a use thereof. The small activating nucleic acid molecule can be a double-stranded or single-stranded RNA molecule targeting the promoter region of the HMBS gene. The first nucleic acid strand and the second nucleic acid strand each contain a complementary region, and the complementary regions can form a double-stranded nucleic acid structure, which can promote the expression of the HMBS gene. The first nucleic acid strand or the second nucleic acid strand independently have a length of 16 to 35 nucleotides. The 3′ terminus of the two oligonucleotide strands may have an overhang of 0 to 6 nucleotides in length. The small activating nucleic acid molecule for the HMBS gene can be used to up-regulate mRNA and protein expressions of the HMBS gene in a cell and promote enzymatic activity thereof

The present disclosure relates to human porphobilinogen deaminase derived proteins and polynucleotides and methods of using these proteins and polynucleotides.

The invention relates to mRNA therapy for the treatment of Acute Intermittent Porphyria (AIP). mRNAs for use in the invention, when administered in vivo, encode human porphobilinogen deaminase (PBGD), isoforms thereof, functional fragments thereof, and fusion proteins comprising PBGD. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to affect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of PBGD expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic metabolites associated with deficient PBGD activity in subjects, namely porphobilinogen and aminolevulinate (PBG and ALA).

The present invention includes a method and kit for detecting porphobilinogen fluorometrically comprising: obtaining a sample suspected of having a porphobilinogen; catalyzing a reaction in the sample with a porphobilinogen deaminase to form a porphyrin; exposing the porphyrin to a light source having a 399 nm wavelength; detecting fluorometrically the amount of porphyrin in the sample; and correlating the amount of porphyrin in the sample to an amount of porphobilinogen in the sample, wherein the ratio of porphobilinogen to porphyrin is 4:1.

The present invention relates to a microorganism variant having the ability to extracellularly produce heme, and more particularly to a metabolically engineered microorganism variant having the ability to extracellularly produce heme and a method of producing heme using the same. According to the present invention, heme, an organometallic compound which is increasingly used as a health food or food supplement for the treatment of porphyria, can be extracellularly secreted and produced in high yield using the microorganism variant, but not conventional chemical synthesis or enzymatic synthesis.