Development of muscular dystrophy in a CRISPR-engineered mutant rabbit model with frame-disrupting ANO5 mutations.
Development of muscular dystrophy in a CRISPR-engineered mutant rabbit model with frame-disrupting ANO5 mutations.
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DOI：
10.1038/s41419-018-0674-y
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发表时间：
2018-05-22
影响因子：
9
通讯作者：
Li Z
中科院分区：
文献类型：
--
作者：
Sui T;Xu L;Lau YS;Liu D;Liu T;Gao Y;Lai L;Han R;Li Z
肢体带状肌营养不良2L型（LGMD2L）和三吉肌病3型（MMD3）是常染色体隐性肌营养不良症，由编码anoctamin-5 （ANO5）的基因突变引起，ANO5属于anoctamin蛋白家族。两个独立的ANO5转录物完全破坏的小鼠系没有表现出明显的肌肉营养不良表型；相反，观察到其中一只小鼠的精子活力出现了一些异常。相比之下，ANO5基因敲除（KO）的第三系小鼠，其ANO5基因缺失表达的残余表现为膜修复缺陷和非常轻微的肌肉病理。许多ANO5相关患者在ANO5基因中携带点突变或小插入/缺失（indels）。为了更接近地模拟人类ANO5突变，我们通过将Cas9 mRNA和sgRNA共同注射到受精卵中来设计ANO5突变兔。在突变家兔中，crispr介导的外显子12和/或13中的小缺失导致肌肉营养不良的典型症状，包括血清肌酸激酶（CK）升高、肌肉坏死、再生、脂肪替代和纤维化。建立ANO5突变兔模型，为ANO5缺乏性肌营养不良的发病机制和治疗方法的研究奠定了基础。
Limb girdle muscular dystrophy type 2L (LGMD2L) and Miyoshi myopathy type 3 (MMD3) are autosomal recessive muscular dystrophy caused by mutations in the gene encoding anoctamin-5 (ANO5), which belongs to the anoctamin protein family. Two independent lines of mice with complete disruption of ANO5 transcripts did not exhibit overt muscular dystrophy phenotypes; instead, one of these mice was observed to present with some abnormality in sperm motility. In contrast, a third line of ANO5-knockout (KO) mice with residual expression of truncated ANO5 expression was reported to display defective membrane repair and very mild muscle pathology. Many of the ANO5-related patients carry point mutations or small insertions/deletions (indels) in the ANO5 gene. To more closely mimic the human ANO5 mutations, we engineered mutant ANO5 rabbits via co-injection of Cas9 mRNA and sgRNA into the zygotes. CRISPR-mediated small indels in the exon 12 and/or 13 in the mutant rabbits lead to the development of typical signs of muscular dystrophy with increased serum creatine kinase (CK), muscle necrosis, regeneration, fatty replacement and fibrosis. This novel ANO5 mutant rabbit model would be useful in studying the disease pathogenesis and therapeutic treatments for ANO5-deficient muscular dystrophy.
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DOI：
10.1016/j.nmd.2015.03.011
发表时间：
2015-07
期刊：
Neuromuscular disorders : NMD
影响因子：
--
作者：
Savarese M;Di Fruscio G;Tasca G;Ruggiero L;Janssens S;De Bleecker J;Delpech M;Musumeci O;Toscano A;Angelini C;Sacconi S;Santoro L;Ricci E;Claes K;Politano L;Nigro V
通讯作者：
Nigro V
影响因子：
9.9
作者：
Penttila, S.;Palmio, J.;Udd, B.
通讯作者：
Udd, B.
影响因子：
3
作者：
Bouquet, F.;Cossee, M.;Eymard, B.
通讯作者：
Eymard, B.
影响因子：
3.7
作者：
Gutpell, Kelly M.;Hrinivich, William T.;Hoffman, Lisa M.
通讯作者：
Hoffman, Lisa M.
影响因子：
3.4
作者：
Little, Ann A.;Mckeever, Paul E.;Gruis, Kirsten L.
通讯作者：
Gruis, Kirsten L.