Abstract: Disclosed are polynucleotides, polypeptides, and gene therapy vectors relating to biologically active methylmalonyl-CoA mutase enzymes, internally tagged with an immunoaffinity and detection epitope, which has been designed and tested in mouse models of methylmalonic acidemia (MMA). The polypeptides and polynucleotides of the present invention contain a mitochondrial leader sequence fused to tag, such as an HA, 3xFLAG, or V5 tag placed in a region of the methylmalonyl-CoA mutase enzyme that maintains mitochondrial localization and function, e.g., the 5? end of a methylmalonyl-CoA mutase polynucleotide is replaced with an engineered nucleotide sequence that encodes the endogenous mitochondrial importation sequence, a mitochondrial protease cleavage site, and a tag. The polynucleotides and polypeptides of the invention are useful to treat conditions such as MMA, and to assay both activity and biodistribution after gene therapy in varied models of MMA.

Abstract: Synthetic polynucleotides encoding human methylmalonyl-CoA mutase (synMUT) and exhibiting augmented expression in cell culture and/or in a subject are described herein. An adeno-associated viral (AAV) gene therapy vector encoding synMUT under the control of a liver-specific promoter (AAV2/8-HCR-hAAT-synMUT-RBG) successfully rescued the neonatal lethal phenotype displayed by methylmalonyl-CoA mutase-deficient mice, lowered circulating methylmalonic acid levels in the treated animals, and resulted in prolonged hepatic expression of the product of synMUT transgene in vivo, human methylmalonyl-CoA mutase (MUT).

Abstract: Synthetic polynucleotides encoding human methylmalonyl-CoA mutase (synMUT) and exhibiting augmented expression in cell culture and/or in a subject are described herein. An adeno-associated viral (AAV) gene therapy vector encoding synMUT under the control of a liver-specific promoter (AAV2/8-HCR-hAAT-synMUT-RBG) successfully rescued the neonatal lethal phenotype displayed by methylmalonyl-CoA mutase-deficient mice, lowered circulating methylmalonic acid levels in the treated animals, and resulted in prolonged hepatic expression of the product of synMUT transgene in vivo, human methylmalonyl-CoA mutase (MUT).

Abstract: Synthetic polynucleotides encoding human methylmalonyl-CoA mutase (synMUT) and exhibiting augmented expression in cell culture and/or in a subject are described herein. An adeno-associated viral (AAV) gene therapy vector encoding synMUT under the control of a liver-specific promoter (AAV2/8-HCR-hAAT-synMUT-RBG) successfully rescued the neonatal lethal phenotype displayed by methylmalonyl-CoA mutase-deficient mice, lowered circulating methylmalonic acid levels in the treated animals, and resulted in prolonged hepatic expression of the product of synMUT transgene in vivo, human methylmalonyl-CoA mutase (MUT).

Abstract: Synthetic polynucleotides encoding human methylmalonyl-CoA mutase (synMUT) and exhibiting augmented expression in cell culture and/or in a subject are described herein. An adeno-associated viral (AAV) gene therapy vector encoding synMUT under the control of a liver-specific promoter (AAV2/8-HCR-hAAT-synMUT-RBG) successfully rescued the neonatal lethal phenotype displayed by methylmalonyl-CoA mutase-deficient mice, lowered circulating methylmalonic acid levels in the treated animals, and resulted in prolonged hepatic expression of the product of synMUT transgene in vivo, human methylmalonyl-CoA mutase (MUT).

Abstract: The invention provides a device including a chamber wherein the chamber including a rigid enclosure; a rigid lid for the enclosure; a gasket between the lid and the enclosure to allow for an airtight seal between the enclosure and the gasket upon closure of a latch connecting the enclosure and the lid; a port for airtight attachment of a syringe, and a port for airtight insertion of a gas sensor. The device can further include a gas sensor and one or more syringes for attachment to the device by a three-way stopcocks. The device is appropriately sized for use with the subject of interest. The invention also provides methods for use of the device.

Abstract: The invention provides a device including a chamber wherein the chamber including a rigid enclosure; a rigid lid for the enclosure; a gasket between the lid and the enclosure to allow for an airtight seal between the enclosure and the gasket upon closure of a latch connecting the enclosure and the lid; a port for airtight attachment of a syringe, and a port for airtight insertion of a gas sensor. The device can further include a gas sensor and one or more syringes for attachment to the device by a three-way stopcocks. The device is appropriately sized for use with the subject of interest. The invention also provides methods for use of the device.

Abstract: The invention provides a device including a chamber wherein the chamber including a rigid enclosure; a rigid lid for the enclosure; a gasket between the lid and the enclosure to allow for an airtight seal between the enclosure and the gasket upon closure of a latch connecting the enclosure and the lid; a port for airtight attachment of a syringe, and a port for airtight insertion of a gas sensor. The device can further include a gas sensor and one or more syringes for attachment to the device by a three-way stopcocks. The device is appropriately sized for use with the subject of interest. The invention also provides methods for use of the device.

Abstract: The invention provides a device including a chamber wherein the chamber including a rigid enclosure; a rigid lid for the enclosure; a gasket between the lid and the enclosure to allow for an airtight seal between the enclosure and the gasket upon closure of a latch connecting the enclosure and the lid; a port for airtight attachment of a syringe, and a port for airtight insertion of a gas sensor. The device can further include a gas sensor and one or more syringes for attachment to the device by a three-way stopcocks. The device is appropriately sized for use with the subject of interest. The invention also provides methods for use of the device.