Abstract: The present disclosure is generally directed to an artificial exonic barcode system. The exonic barcodes comprise a nucleotide sequence comprising from 5? to 3? a 5? barcode, an intron, and a 3? barcode, and the disclosure is further directed to a library of these exonic barcodes. The disclosure also describes a method of generating the exonic barcode library and using the library of exonic barcodes in a method of screening for efficiency of transformation and/or expression of one or more genetic constructs in a subject. Primers and probes were also designed for validation of these exonic barcodes and corresponding methods.

Abstract: Disclosed herein are methods for systemically editing a gene in a subject and for systemically treating a genetic condition in a subject using a dual-vector CRISPR-Cas therapy. The methods comprise administering to the subject, via systemic administration, a gene editing AAV vector encoding a CRISPR effector protein (e.g., a Cas protein) and a targeting AAV vector providing one or more gRNAs targeted to the gene. In the methods, the ratio of the targeting AAV vector to the gene editing vector is greater than or equal to 2. Also provided are dual-vector systems for editing a gene or treating a genetic disease in a subject.

Abstract: Synthetic nucleic acids encoding mini and microdystrophin genes comprising the membrane binding motifs or domains of the R10-R11-R12 region are provided. Also provided are vectors, host cells, and related methods of using the same to treat a subject suffering from Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) or X-linked dilated cardiomyopathy (XLDC), or for ameliorating one or more adverse effects of DMD, BMD, or XLDC. Also provided are a fusion protein comprising a nNOS binding domain of dystrophin R16-R17 that is operably linked to a syntrophin PDZ domain and synthetic nucleic acids comprising the same that can be used to treat subjects with diseases characterized by loss of sarcolemmal neuronal nitric oxide synthase (nNOS) activity.

Abstract: Disclosed are recombinant adeno-associated virus (rAAV) nucleic acid vectors comprising inverted terminal repeats (ITRs) free of 5?-cytosine-phosphate-guanine-3? (CpG) motifs. Also disclosed are rAAV particles comprising the rAAV vectors, and to compositions and methods for delivering nucleic acids and/or for gene therapy. Further disclosed are compositions and methods for treating diseases with AAV gene therapy using the rAAV vector.

Abstract: Synthetic nucleic acids encoding mini and microdystrophin genes comprising the membrane binding motifs or domains of the R10-R11-R12 region are provided. Also provided are vectors, host cells, and related methods of using the same to treat a subject suffering from Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) or X-linked dilated cardiomyopathy (XLDC), or for ameliorating one or more adverse effects of DMD, BMD, or XLDC. Also provided are a fusion protein comprising a nNOS binding domain of dystrophin R16-R17 that is operably linked to a syntrophin PDZ domain and synthetic nucleic acids comprising the same that can be used to treat subjects with diseases characterized by loss of sarcolemmal neuronal nitric oxide synthase (nNOS) activity.

Abstract: High-resolution 3D optical polarization tractography (OPT) images of the internal fiber structure of a target tissue. Manipulation of dual-angle imaging data of the fiber orientation inside a target tissue leads to the determination of 3D imaging properties of the target tissue, allowing transmission of the 3D image properties of the target tissue to an OPT processor to produce high-resolution 3D images.

Abstract: Synthetic nucleic acids encoding mini and microdystrophin genes comprising the membrane binding motifs or domains of the R10-R11-R12 region are provided. Also provided are vectors, host cells, and related methods of using the same to treat a subject suffering from Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) or X-linked dilated cardiomyopathy (XLDC), or for ameliorating one or more adverse effects of DMD, BMD, or XLDC. Also provided are a fusion protein comprising a nNOS binding domain of dystrophin R16-R17 that is operably linked to a syntrophin PDZ domain and synthetic nucleic acids comprising the same that can be used to treat subjects with diseases characterized by loss of sarcolemmal neuronal nitric oxide synthase (nNOS) activity.

Abstract: Disclosed herein are methods for systemically editing a gene in a subject and for systemically treating a genetic condition in a subject using a dual-vector CRISPR-Cas therapy. The methods comprise administering to the subject, via systemic administration, a gene editing AAV vector encoding a CRISPR effector protein (e.g., a Cas protein) and a targeting AAV vector providing one or more gRNAs targeted to the gene. In the methods, the ratio of the targeting AAV vector to the gene editing vector is greater than or equal to 2. Also provided are dual-vector systems for editing a gene or treating a genetic disease in a subject.

Abstract: Disclosed are compositions and methods for treating dystrophinopathies. Compositions include modified dystrophin polynucleotides that encode modified dystrophin proteins having modified hinge 1 (H1) and/or hinge 4 (H4). Also disclosed are methods for treating dystrophinopathies by administering compositions encoding modified dystrophin proteins having modified hinge 1 (H1) and/or modified hinge 4 (H4).

Abstract: According to the embodiments described herein, a series of biological materials for treatment/therapy of DMD and/or BMD through the recovery of sarcolemmal nNOS is provided. The biological material comprises the complete dystrophin repeats R16 and R17 or certain domains, sections, or fragments of the dystrophin repeats R16 and R17. In some aspects, such domains, sections, or fragments may be selected from sequence motifs including dystrophin R17 ?1 helix, ?2 and ?3 helices of both R16 and R17, or a combination thereof.

Abstract: Synthetic nucleic acids encoding mini and microdystrophin genes comprising the membrane binding motifs or domains of the R10-R11-R12 region are provided. Also provided are vectors, host cells, and related methods of using the same to treat a subject suffering from Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) or X-linked dilated cardiomyopathy (XLDC), or for ameliorating one or more adverse effects of DMD, BMD, or XLDC. Also provided are a fusion protein comprising a nNOS binding domain of dystrophin R16-R17 that is operably linked to a syntrophin PDZ domain and synthetic nucleic acids comprising the same that can be used to treat subjects with diseases characterized by loss of sarcolemmal neuronal nitric oxide synthase (nNOS) activity.

Abstract: High-resolution 3D optical polarization tractography (OPT) images of the internal fiber structure of a target tissue. Manipulation of dual-angle imaging data of the fiber orientation inside a target tissue leads to the determination of 3D imaging properties of the target tissue, allowing transmission of the 3D image properties of the target tissue to an OPT processor to produce high-resolution 3D images.

Abstract: High-resolution 3D optical polarization tractography (OPT) images of the internal fiber structure of a target tissue. Manipulation of dual-angle imaging data of the fiber orientation inside a target tissue leads to the determination of 3D imaging properties of the target tissue, allowing transmission of the 3D image properties of the target tissue to an OPT processor to produce high-resolution 3D images.

Abstract: According to the embodiments described herein, a series of biological materials for treatment/therapy of DMD and/or BMD through the recovery of sarcolemmal nNOS is provided. The biological material comprises the complete dystrophin repeats R16 and R17 or certain domains, sections, or fragments of the dystrophin repeats R16 and R17. In some aspects, such domains, sections, or fragments may be selected from sequence motifs including dystrophin R17 ?1 helix, ?2 and ?3 helices of both R16 and R17, or a combination thereof.

Abstract: According to the embodiments described herein, a series of biological materials for treatment/therapy of DMD and/or BMD through the recovery of sarcolemmal nNOS is provided. The biological material comprises the complete dystrophin repeats R16 and R17 or certain domains, sections, or fragments of the dystrophin repeats R16 and R17. In some aspects, such domains, sections, or fragments may be selected from sequence motifs including dystrophin R17 ?1 helix, ?2 and ?3 helices of both R16 and R17, or a combination thereof.

Abstract: High-resolution 3D optical polarization tractography (OPT) images of the internal fiber structure of a target tissue. Manipulation of dual-angle imaging data of the fiber orientation inside a target tissue leads to the determination of 3D imaging properties of the target tissue, allowing transmission of the 3D image properties of the target tissue to an OPT processor to produce high-resolution 3D images.

Abstract: Agents and methods to alter rAAV transduction are provided.

Abstract: According to the embodiments described herein, a series of biological materials for treatment/therapy of DMD and/or BMD through the recovery of sarcolemmal nNOS is provided. The biological material comprises the complete dystrophin repeats R16 and R17 or certain domains, sections, or fragments of the dystrophin repeats R16 and R17. In some aspects, such domains, sections, or fragments may be selected from sequence motifs including dystrophin R17 ?1 helix, ?2 and ?3 helices of both R16 and R17, or a combination thereof.

Abstract: The present invention provides novel dystrophin mini/micro-genes that retain the essential biological functions of a full-length dystrophin gene. More particularly, the present invention provides to a series of synthetic mini/micro-dystrophin genes capable of restoring neuronal nitric oxide synthase (nNOS) to the sarcolemma. A method as well as a pharmaceutical composition for treatment of Duchenne Muscular Dystrophy (DMD), Becker Muscular Dystrophy (BMD), and X-linked Dilated Cardiomyopathy (XLDC) are also provided.

Abstract: The invention provides an isolated and purified DNA molecule comprising at least one DNA segment, a biologically active subunit or variant thereof, of a circular intermediate of adeno-associated virus, which DNA segment confers increased episomal stability, persistence or abundance of the isolated DNA molecule in a host cell. The invention also provides a composition comprising at least two adeno-associated virus vectors.