Quantitative evaluation of incomplete preweaning lethality in mice by using the CRISPR/Cas9 system.
Quantitative evaluation of incomplete preweaning lethality in mice by using the CRISPR/Cas9 system.
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DOI：
10.1038/s41598-018-34270-5
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发表时间：
2018-10-30
影响因子：
4.6
通讯作者：
Kato T
中科院分区：
文献类型：
--
作者：
Nakamura T;Nakajima K;Ohnishi T;Yoshikawa T;Nakanishi M;Takumi T;Tsuboi T;Kato T
实施基因组编辑的各种分子生物学技术使得产生几乎所有已知基因的小鼠突变体成为可能。由此，建立了国际小鼠表型联盟（IMPC）数据库，其中列出了转基因小鼠的表型。在小鼠表型中，致死率对于评估基因在小鼠生存中的重要性至关重要。尽管据报道许多基因在IMPC数据库中显示出“断奶前致死率、不完全外显率”，但纯合基因敲除小鼠的存活率差异很大。在这里，我们提出致死等位基因指数（LAI），即观察到的纯合基因敲除（KO）小鼠数量与理论预测纯合KO小鼠数量的比率，作为断奶前致死率的简单定量指标。在 IMPC 中登记为不完全致死的小鼠突变体中，根据 F1 小鼠基因型计算的 LAI 在疾病相关基因中往往较低，并且与人类功能丧失 (LOF) 等位基因的频率相关。在使用 CRISPR/Cas9 进行基因组编辑的小鼠中，具有纯合移码等位基因的小鼠数量似乎与致死率相关。我们使用CRISPR/Cas9编辑了细胞系和小鼠中的Ehd1基因，发现基因型分布存在显着差异。根据这些数据计算的 LAI 与根据 IMPC 数据计算的值相似。这些发现支持 LAI 作为基因组编辑小鼠断奶前致死率指标的潜在用途。
Various molecular biology techniques implementing genome editing have made it possible to generate mouse mutants for nearly all known genes; as a result, the International Mouse Phenotyping Consortium (IMPC) database listing the phenotypes of genetically modified mice has been established. Among mouse phenotypes, lethality is crucial to evaluate the importance of genes in mouse survival. Although many genes are reported to show “preweaning lethality, incomplete penetrance” in the IMPC database, the survival rates of homozygous knockout mice are highly variable. Here, we propose the lethal allele index (LAI), the ratio of the observed number of mice with homozygous knockout (KO) to the theoretically predicted number of homozygous KO mice, as a simple quantitative indicator of preweaning lethality. Among the mice mutants registered as incompletely lethal in IMPC, the LAI calculated from the genotypes of F1 mice tended to be lower in disease-related genes, and correlated with the frequency of loss-of-function (LOF) alleles in humans. In genome-edited mice using CRISPR/Cas9, the number of mice with homozygous frameshift alleles seemed to be associated with lethality. We edited the Ehd1 gene in cell lines as well as mice using CRISPR/Cas9, and found that the genotype distribution was significantly different. The LAI calculated from these data was similar to the value calculated from the IMPC data. These findings support the potential usefulness of the LAI as an index of preweaning lethality in genome-edited mice.
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影响因子：
30.8
作者：
Meehan TF;Conte N;West DB;Jacobsen JO;Mason J;Warren J;Chen CK;Tudose I;Relac M;Matthews P;Karp N;Santos L;Fiegel T;Ring N;Westerberg H;Greenaway S;Sneddon D;Morgan H;Codner GF;Stewart ME;Brown J;Horner N;International Mouse Phenotyping Consortium;Haendel M;Washington N;Mungall CJ;Reynolds CL;Gallegos J;Gailus-Durner V;Sorg T;Pavlovic G;Bower LR;Moore M;Morse I;Gao X;Tocchini-Valentini GP;Obata Y;Cho SY;Seong JK;Seavitt J;Beaudet AL;Dickinson ME;Herault Y;Wurst W;de Angelis MH;Lloyd KCK;Flenniken AM;Nutter LMJ;Newbigging S;McKerlie C;Justice MJ;Murray SA;Svenson KL;Braun RE;White JK;Bradley A;Flicek P;Wells S;Skarnes WC;Adams DJ;Parkinson H;Mallon AM;Brown SDM;Smedley D
通讯作者：
Smedley D
通过 CRISPR/Cas9 介导的同源独立靶向整合进行体内基因组编辑
影响因子：
64.8
作者：
Suzuki K;Tsunekawa Y;Hernandez-Benitez R;Wu J;Zhu J;Kim EJ;Hatanaka F;Yamamoto M;Araoka T;Li Z;Kurita M;Hishida T;Li M;Aizawa E;Guo S;Chen S;Goebl A;Soligalla RD;Qu J;Jiang T;Fu X;Jafari M;Esteban CR;Berggren WT;Lajara J;Nuñez-Delicado E;Guillen P;Campistol JM;Matsuzaki F;Liu GH;Magistretti P;Zhang K;Callaway EM;Zhang K;Belmonte JC
通讯作者：
Belmonte JC
影响因子：
4.6
作者：
Nakajima K;Kazuno AA;Kelsoe J;Nakanishi M;Takumi T;Kato T
通讯作者：
Kato T
DOI：
10.4049/jimmunol.1601793
发表时间：
2018-01-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Iseka FM;Goetz BT;Mushtaq I;An W;Cypher LR;Bielecki TA;Tom EC;Arya P;Bhattacharyya S;Storck MD;Semerad CL;Talmadge JE;Mosley RL;Band V;Band H
通讯作者：
Band H
影响因子：
21.3
作者：
Wu, Chien-Ting;Chen, Hsin-Yi;Tang, Tang K.
通讯作者：
Tang, Tang K.