Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish.
Reverse genetic screening reveals poor correlation between morpholino-induced and mutant phenotypes in zebrafish.
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DOI：
10.1016/j.devcel.2014.11.018
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发表时间：
2015-01-12
影响因子：
11.8
通讯作者：
Lawson, Nathan D.
中科院分区：
文献类型：
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作者：
Kok, Fatma O.;Shin, Masahiro;Ni, Chih-Wen;Gupta, Ankit;Grosse, Ann S.;van Impel, Andreas;Kirchmaier, Bettina C.;Peterson-Maduro, Josi;Kourkoulis, George;Male, Ira;DeSantis, Dana F.;Sheppard-Tindell, Sarah;Ebarasi, Lwaki;Betsholtz, Christer;Schulte-Merker, Stefan;Wolfe, Scot A.;Lawson, Nathan D.
可编程位点特异性核酸酶的广泛应用现在可以实现斑马鱼的靶向基因破坏。在这项研究中，我们应用位点特异性核酸酶来生成在二十多个基因中带有单个突变的斑马鱼品系。我们发现只有一小部分基因的突变导致了胚胎发生的缺陷。此外，十种不同基因的突变体未能重现已发表的吗啡啉诱导的表型（形态）。突变胚胎中表型的缺失不太可能是由于母体效应或未能消除基因功能所致。一致地，将已发表的 morphant 缺陷与桑格斑马鱼突变项目进行比较表明，大约 80% 的 morphant 表型在突变体胚胎中未观察到，与我们的突变体收集相似。基于这些结果，我们建议突变表型成为定义斑马鱼基因功能的标准指标，之后重演各自表型的吗啡啉可以可靠地应用于辅助分析。
The widespread availability of programmable site-specific nucleases now enables targeted gene disruption in the zebrafish. In this study, we applied site-specific nucleases to generate zebrafish lines bearing individual mutations in more than twenty genes. We found that mutations in only a small proportion of genes caused defects in embryogenesis. Moreover, mutants for ten different genes failed to recapitulate published Morpholino-induced phenotypes (morphants). The absence of phenotypes in mutant embryos was not likely due to maternal effects or failure to eliminate gene function. Consistently, a comparison of published morphant defects with the Sanger Zebrafish Mutation Project revealed that approximately eighty percent of morphant phenotypes were not observed in mutant embryos, similar to our mutant collection. Based on these results, we suggest that mutant phenotypes become the standard metric to define gene function in zebrafish, after which Morpholinos that recapitulate respective phenotypes could be reliably applied for ancillary analyses.
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影响因子：
3.7
作者：
Chapman AL;Bennett EJ;Ramesh TM;De Vos KJ;Grierson AJ
通讯作者：
Grierson AJ
影响因子：
14.9
作者：
Cade L;Reyon D;Hwang WY;Tsai SQ;Patel S;Khayter C;Joung JK;Sander JD;Peterson RT;Yeh JR
通讯作者：
Yeh JR
影响因子：
1.5
作者：
Draper, BW;Morcos, PA;Kimmel, CB
通讯作者：
Kimmel, CB
DOI：
10.1073/pnas.0506886103
发表时间：
2006-04-25
影响因子：
11.1
作者：
Covassin, LD;Villefranc, JA;Lawson, ND
通讯作者：
Lawson, ND
影响因子：
64.8
作者：
通讯作者：
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