Insertion of a knockout-first cassette in Ampd1 gene leads to neonatal death by disruption of neighboring genes expression.
Insertion of a knockout-first cassette in Ampd1 gene leads to neonatal death by disruption of neighboring genes expression.
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在 Ampd1 基因中插入先敲除盒会破坏邻近基因的表达，从而导致新生儿死亡。
DOI：
10.1038/srep35970
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发表时间：
2016-10-24
影响因子：
4.6
通讯作者：
Li JD
中科院分区：
文献类型：
--
作者：
Pan Y;Zhang L;Liu Q;Li Y;Guo H;Peng Y;Peng H;Tang B;Hu Z;Zhao J;Xia K;Li JD
AMPD1是一种单磷酸腺苷脱氨酶，催化AMP脱氨为IMP。为了了解AMPD1的生理功能，我们从KOMP库中获得了一株Ampd1突变小鼠，该突变小鼠是通过基因敲除优先策略产生的。E18.5Ampd1tm1a/tm1a小鼠骨骼肌AMP水平升高，IMP几乎完全缺失。然而，Ampd1tm1a/tm1a小鼠在出生后2天内死亡，这与以前的报道相矛盾。在移除了第一个敲除的盒式磁带和关键外显子后，Ampd1tm1c/tm1c和Ampd1tm1d/tm1d等位基因纯合的小鼠存活到成年。RNA-seq分析表明，两个相邻基因Man1a2和NRAS在Ampd1tm1a/tm1a小鼠中表达中断，而在Ampd1tm1c/tm1c和Ampd1tm1d/tm1d小鼠中正常表达。Ampd1tm1a/tm1a小鼠的新生儿致死性表型与Man1a2缺陷小鼠一致。我们的结果表明，基因敲除第一盒可能通过影响邻近基因的表达而引起脱靶效应。这项研究与其他报告一起强烈表明，通过位点特异性重组酶去除靶向盒对于靶标突变产生的小鼠的准确表型解释非常重要。
AMPD1 is an adenosine monophosphate deaminase that catalyzes the deamination of AMP to IMP. To understand the physiological function of AMPD1, we obtained a strain of Ampd1 mutant mice from KOMP repository, which was generated by a knockout-first strategy. An elevated AMP level and almost complete lack of IMP was detected in the skeletal muscle of E18.5 Ampd1tm1a/tm1a mice. However, Ampd1tm1a/tm1a mice died in 2 days postnatally, which was contradicting to previous reports. After removal of the knockout-first cassette and critical exon, mice homozygous for the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d alleles survived to adulthood. RNA-seq analysis indicated that the expression of two neighboring genes, Man1a2 and Nras, were disrupted in the Ampd1tm1a/tm1a mice, but normal in the Ampd1tm1c/tm1c and Ampd1tm1d/tm1d mice. The neonatal lethality phenotype in the Ampd1tm1a/tm1a mice was consistent with the Man1a2-deficient mice. Our results indicated the knockout-first cassette may cause off-target effect by influence the expression of neighboring genes. This study, together with other reports, strongly suggests that removal of targeting cassette by site-specific recombinases is very important for the accurate phenotypic interpretation on mice generated by target mutations.
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影响因子：
64.8
作者：
Skarnes WC;Rosen B;West AP;Koutsourakis M;Bushell W;Iyer V;Mujica AO;Thomas M;Harrow J;Cox T;Jackson D;Severin J;Biggs P;Fu J;Nefedov M;de Jong PJ;Stewart AF;Bradley A
通讯作者：
Bradley A
影响因子：
--
作者：
Plaideau, Catheline;Lai, Yu-Chiang;Rider, Mark H.
通讯作者：
Rider, Mark H.
1p13.2 上的常见遗传变异与自闭症风险相关。
影响因子：
11
作者：
Xia, K.;Guo, H.;Zhang, F.
通讯作者：
Zhang, F.
影响因子：
2.6
作者：
KOSTIC, D;CAPECCHI, MR
通讯作者：
CAPECCHI, MR
影响因子：
10.5
作者：
FIERING, S;EPNER, E;GROUDINE, M
通讯作者：
GROUDINE, M