Unintentional miRNA ablation is a risk factor in gene knockout studies: a short report.
Unintentional miRNA ablation is a risk factor in gene knockout studies: a short report.
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无意的miRNA消融是基因敲除研究的危险因素：简短的报告。
DOI：
10.1371/journal.pgen.0040034
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发表时间：
2008-02
期刊：
影响因子：
4.5
通讯作者：
McManus, Michael T.
中科院分区：
文献类型：
--
作者：
Osokine, Ivan;Hsu, Ruby;Loeb, Gabriel B.;McManus, Michael T.
用于研究基因功能的最有力的技术之一是使用基因工程策略（例如靶向重组或基因捕获盒的病毒整合）破坏该基因的表达。这些工具的巨大实用性在今年获得了诺贝尔生理学或医学奖，以表彰Capecchi，Evans和Smithies在哺乳动物靶向重组诱变方面的开创性工作。另一个值得注意的发现是近十年前发现了一类称为microRNA的新型非编码基因。MicroRNA是已知最大的调控元件类别之一，预计在小鼠基因组中存在超过1000种。超过50%的已知microRNA位于编码基因的内含子内。鉴于目前小鼠中约有一半的基因已被敲除，我们研究了内含子microRNA在其中一些小鼠模型中可能被巧合地删除或破坏的可能性。我们检索了已发表的小鼠基因敲除研究和基因陷阱胚胎干细胞系数据库，以寻找microRNA位于操纵基因组位点内或附近的病例，发现近200例microRNA表达可能与另一个基因沿着被破坏。我们的研究结果引起了人们的注意，需要在未来的敲除研究中仔细规划，以尽量减少microRNA的无意破坏。这些数据也提出了许多敲除研究可能需要重新检查的可能性，以确定是否microRNA的丢失有助于归因于蛋白质编码基因丢失的表型后果。为了确定基因的功能，通常首先通过靶向重组或插入基因捕获盒来破坏该基因的表达。在我们的研究中，我们指出这些方法可能会被称为microRNA的小非编码元件的存在所混淆。microRNA构成了最大类的调控元件之一，并且超过50%的已知microRNA已经在编码基因的内含子内被鉴定。因此，基因的破坏也可能导致该区域微小RNA的破坏。在这项研究中，我们搜索了基因捕获细胞系的数据库以及先前发表的敲除研究，并报告了近200个microRNA表达可能被无意中断的例子。我们的研究结果具有广泛的兴趣和重要性，因为它们提出了可能需要重新检查一些蛋白质功能研究的可能性，以确定microRNA的丢失是否可能导致先前归因于蛋白质丢失的表型。
One of the most powerful techniques for studying the function of a gene is to disrupt the expression of that gene using genetic engineering strategies such as targeted recombination or viral integration of gene trap cassettes. The tremendous utility of these tools was recognized this year with the awarding of the Nobel Prize in Physiology or Medicine to Capecchi, Evans, and Smithies for their pioneering work in targeted recombination mutagenesis in mammals. Another noteworthy discovery made nearly a decade ago was the identification of a novel class of non-coding genes called microRNAs. MicroRNAs are among the largest known classes of regulatory elements with more than 1000 predicted to exist in the mouse genome. Over 50% of known microRNAs are located within introns of coding genes. Given that currently about half of the genes in mouse have been knocked out, we investigated the possibility that intronic microRNAs may have been coincidentally deleted or disrupted in some of these mouse models. We searched published murine knockout studies and gene trap embryonic stem cell line databases for cases where a microRNA was located within or near the manipulated genomic loci, finding almost 200 cases where microRNA expression may have been disrupted along with another gene. Our results draw attention to the need for careful planning in future knockout studies to minimize the unintentional disruption of microRNAs. These data also raise the possibility that many knockout studies may need to be reexamined to determine if loss of a microRNA contributes to the phenotypic consequences attributed to loss of a protein-encoding gene. To determine the function of a gene, it is often informative to first disrupt the expression of that gene through targeted recombination or the insertion of gene trap cassettes. In our study, we point out that these approaches may be confounded by the presence of small non-coding elements known as microRNAs. MicroRNAs constitute one of the largest classes of regulatory elements, and over 50% of known microRNAs have been identified within an intron of a coding gene. Disruption of a gene could therefore also result in the disruption of microRNAs in the region. In this study, we searched databases of gene-trapped cell lines as well as previously published knockout studies and report almost 200 examples where microRNA expression may have been unintentionally disrupted. Our results are of broad interest and importance because they raise the possibility that a number of protein function studies may need to be reexamined to determine whether the loss of a microRNA may have contributed to the phenotype previously attributed to the loss of a protein.
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影响因子：
2.6
作者：
Liu, JM;Zhang, L;Hu, HY
通讯作者：
Hu, HY
影响因子：
14.9
作者：
Griffiths-Jones S;Grocock RJ;van Dongen S;Bateman A;Enright AJ
通讯作者：
Enright AJ
影响因子：
7
作者：
Rodriguez, A;Griffiths-Jones, S;Bradley, A
通讯作者：
Bradley, A
影响因子：
14.9
作者：
Eppig JT;Bult CJ;Kadin JA;Richardson JE;Blake JA;Anagnostopoulos A;Baldarelli RM;Baya M;Beal JS;Bello SM;Boddy WJ;Bradt DW;Burkart DL;Butler NE;Campbell J;Cassell MA;Corbani LE;Cousins SL;Dahmen DJ;Dene H;Diehl AD;Drabkin HJ;Frazer KS;Frost P;Glass LH;Goldsmith CW;Grant PL;Lennon-Pierce M;Lewis J;Lu I;Maltais LJ;McAndrews-Hill M;McClellan L;Miers DB;Miller LA;Ni L;Ormsby JE;Qi D;Reddy TB;Reed DJ;Richards-Smith B;Shaw DR;Sinclair R;Smith CL;Szauter P;Walker MB;Walton DO;Washburn LL;Witham IT;Zhu Y;Mouse Genome Database Group
通讯作者：
Mouse Genome Database Group