Assisted reproduction mediated resurrection of a feline model for Chediak-Higashi syndrome caused by a large duplication in LYST
Assisted reproduction mediated resurrection of a feline model for Chediak-Higashi syndrome caused by a large duplication in LYST
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DOI：
10.1038/s41598-019-56896-9
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发表时间：
2020-01-09
影响因子：
4.6
通讯作者：
Lyons, L. A.
中科院分区：
文献类型：
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作者：
Buckley, R. M.;Grahn, R. A.;Lyons, L. A.
Chediak-Higashi综合征(CHS)是一种由溶酶体转运调节因子(Lyst)突变引起的常染色体隐性遗传性溶酶体疾病。CHS的猫科动物模型最初被维持了大约20年。然而，在确定致病突变之前，菌落被解散，CHS猫模型被研究社区丢失。为了复活猫模型，从一名孤独的、有生育能力的CHS携带者身上收集并冷冻精液。用冷冻精液对3只母猫进行了腹腔镜输卵管人工授精，其中2只母猫产下了11只存活的小猫。为了确定致病突变，对来自原始群体的一只受影响的猫的成纤维细胞系进行了全基因组测序。对序列数据的目测检查发现，候选原因变体类似于Lyst内的20kb串联复制，跨越外显子30至38(NM_001290242.1：C.8347-24229548+1749dup)。对产生的后代进行的PCR基因分型显示，有三个人遗传了CHS携带者男性的突变等位基因。本研究成功地利用冷冻保存和辅助生殖来维持和复活生物医学模型，并确定了导致家猫Chediak-Higashi综合征的变异。
Chediak-Higashi Syndrome (CHS) is a well-characterized, autosomal recessively inherited lysosomal disease caused by mutations in lysosomal trafficking regulator (LYST). The feline model for CHS was originally maintained for similar to 20 years. However, the colonies were disbanded and the CHS cat model was lost to the research community before the causative mutation was identified. To resurrect the cat model, semen was collected and cryopreserved from a lone, fertile, CHS carrier male. Using cryopreserved semen, laparoscopic oviductal artificial insemination was performed on three queens, two queens produced 11 viable kittens. To identify the causative mutation, a fibroblast cell line, derived from an affected cat from the original colony, was whole genome sequenced. Visual inspection of the sequence data identified a candidate causal variant as a similar to 20 kb tandem duplication within LYST, spanning exons 30 through to 38 (NM_001290242.1:c.8347-2422_9548 + 1749dup). PCR genotyping of the produced offspring demonstrated three individuals inherited the mutant allele from the CHS carrier male. This study demonstrated the successful use of cryopreservation and assisted reproduction to maintain and resurrect biomedical models and has defined the variant causing Chediak-Higashi syndrome in the domestic cat.