公开(公告)号：US20220072156A1

公开(公告)日：2022-03-10

申请号：US17404915

申请日：2021-08-17

Applicant: The Board of Regents of the University of Texas System

Inventor： Eric N. OLSON ,  Chengzu LONG ,  John R. McANALLY ,  John M. SHELTON ,  Rhonda BASSEL-DUBY

IPC: A61K48/00 ,  A61K38/46 ,  C12N15/113

Abstract: Duchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. The disclosure reports CRISPR/Cas9-mediated gene editing (Myo-editing) is effective at correcting the dystrophin gene mutation in the mdx mice, a model for DMD. Further, the disclosure reports optimization of germline editing of mdx mice by engineering the permanent skipping of mutant exon (exon 23) and extending exon skipping to also correct the disease by post-natal delivery of adeno-associate virus (AAV). AAV-mediated Myo-editing can efficiently rescue the reading frame of dystrophin in mdx mice in vivo. The disclosure reports means of Myo-editing-mediated exon skipping has been successfully advanced from somatic tissues in mice to human DMD patients-derived iPSCs (induced pluripotent stem cells). Custom Myo-editing was performed on iPSCs from patients with differing mutations and successfully restored dystrophin protein expression for all mutations in iPSCs-derived cardiomyocytes.

公开(公告)号：US20160058889A1

公开(公告)日：2016-03-03

申请号：US14823563

申请日：2015-08-11

Applicant: The Board of Regents of the University of Texas System

Inventor： Eric N. OLSON ,  Chengzu LONG ,  John R. MCANALLY ,  John M. SHELTON ,  Rhonda BASSEL-DUBY

IPC: A61K48/00 ,  C12N15/113 ,  A61K38/46

CPC classification number: A61K48/0058 ,  A61K38/465 ,  A61P21/00 ,  C12N15/113 ,  C12N2310/20 ,  C12N2750/14143 ,  C12Y301/00

Abstract: Duchenne muscular dystrophy (DMD) is an inherited X-linked disease caused by mutations in the gene encoding dystrophin, a protein required for muscle fiber integrity. The disclosure reports CRISPR/Cas9-mediated gene editing (Myo-editing) is effective at correcting the dystrophin gene mutation in the mdx mice, a model for DMD. Further, the disclosure reports optimization of germline editing of mdx mice by engineering the permanent skipping of mutant exon (exon 23) and extending exon skipping to also correct the disease by post-natal delivery of adeno-associate virus (AAV). AAV-mediated Myo-editing can efficiently rescue the reading frame of dystrophin in mdx mice in vivo. The disclosure reports means of Myo-editing-mediated exon skipping has been successfully advanced from somatic tissues in mice to human DMD patients-derived iPSCs (induced pluripotent stem cells). Custom Myo-editing was performed on iPSCs from patients with differing mutations and successfully restored dystrophin protein expression for all mutations in iPSCs-derived cardiomyocytes.

Abstract translation: 杜氏肌营养不良（DMD）是由编码肌营养不良蛋白的基因（肌纤维完整性所需的蛋白质）的突变引起的遗传性X连锁疾病。 披露报告CRISPR / Cas9介导的基因编辑（Myo-editing）在校正mdx小鼠中的肌营养不良蛋白基因突变中是有效的，DMD的模型。 此外，该公开报告通过工程化突变外显子（外显子23）的永久性跳过并延伸外显子跳过以通过产后输送腺相关病毒（AAV）来纠正疾病来报告mdx小鼠的种系编辑的优化。 AAV介导的Myo编辑可以有效地挽救体内mdx小鼠肌营养不良蛋白的阅读框架。 Myo编辑介导的外显子跳跃的披露报告手段已经从小鼠的体细胞组织中成功推进到人DMD患者来源的iPSC（诱导多能干细胞）。 在具有不同突变的患者的iPSC上进行定制Myo编辑，并成功地恢复了iPSC来源的心肌细胞中所有突变的肌营养不良蛋白表达。