Intersectional gene inactivation: there is more to conditional mutagenesis than Cre.
Intersectional gene inactivation: there is more to conditional mutagenesis than Cre.
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DOI：
10.1007/s11427-018-9291-2
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发表时间：
2018-09
期刊：
影响因子：
--
通讯作者：
Soriano P
中科院分区：
文献类型：
--
作者：
Soriano P
全世界许多实验室和大规模国际诱变联合体的努力已经导致几乎所有小鼠基因的诱变，并且目前的努力致力于种系无效突变小鼠的大规模表型分析（Ayadi et al.，2012年）。尽管与给定基因的功能丧失相关的早期致死性可能阻止阐明其在后期或在给定组织中的功能，但其中基因的关键部分侧翼为重组酶靶位点的条件等位基因为解决后期基因功能提供了机会（Branda和Dymecki，2004）。幸运的是，由聚生体产生的许多突变等位基因允许使用定点重组酶进行条件诱变。多年来，用于此目的的最广泛使用的重组酶是来自噬菌体P1的Cre重组酶。Cre重组酶催化以正向方向侧接loxP位点的序列的切除，或以相反方向侧接loxP位点的序列的倒位。此外，可调节Cre融合的可用性允许通过外源性他莫昔芬或孕酮施用来调节酶活性，这是在发育后期发生谱系定型时的有用特征。因此，大量Cre驱动系是可用的，如果使用诱导型Cre，则其提供组织特异性或时间控制（http：//www.cre.信息学. Jax. org/home/recombinase）。不幸的是，这些线可能并不总是提供精确的组织特异性，需要阐明突变体表型。作为从我们自己的研究中取得的一个例子，在颅面发育领域工作的许多研究人员依赖于Wnt 1Cre或Wnt 1Cre 2驱动器来去除整个神经嵴的基因功能，这产生了大部分面部骨骼（Danielian等人，1998;刘易斯等人，2013年）。虽然这些线非常有用，但不足以解决与神经嵴或颅面发育中基因功能相关的所有问题。Wnt 1Cree/Wnt 1Cre 2在神经嵴细胞被指定后开启（Barriga等，2015），因此这些品系无法解决与所研究的基因在神经嵴特化中的可能作用有关的问题。Wnt 1Cre/Wnt 1Cre 2也非常广泛地标记神经嵴细胞，因此不能解决特定基因在关键颅面亚区中的作用，所述关键颅面亚区可导致面部裂开表型，例如我们用Pdgfra或Fgfr 1 Wnt 1Cre条件突变体观察到的那些（Brewer et al.，2015; Tallquist和Soriano，
The efforts of many laboratories worldwide and large scale international mutagenesis consortia have resulted in mutagenesis of nearly all mouse genes, and present efforts are devoted to large scale phenotyping of germline null mutant mice (Ayadi et al., 2012). Although early lethality associated with loss of function of a given gene may prevent elucidating its function at a later stage or in a given tissue, conditional alleles in which a critical part of the gene is flanked by recombinase target sites provide an opportunity for addressing gene function at later stages (Branda and Dymecki, 2004). Fortunately, many of the mutant alleles generated by the consortia allow for conditional mutagenesis using site directed recombinases.Over the years, the most widely used recombinase used for this purpose has been the Cre recombinase from the bacteriophage P1. Cre recombinase catalyzes the excision of sequences flanked by loxP sites in direct orientation, or the inversion of sequences flanked by loxP sites in opposite orientation. The availability of regulatable Cre fusions moreover allows enzymatic activity to be regulated by exogenous tamoxifen or progesterone administration, a useful feature when lineage commitment takes place at later stages of development. A large collection of Cre driver lines is thus available that provide tissue specificity or temporal control if using an inducible Cre (http://www. informatics. jax. org/home/recombinase). Unfortunately, these lines may not always offer the precise tissue specificity that is required to elucidate a mutant phenotype. As one example taken from our own line of research, many investigators working in the field of craniofacial development have relied on the Wnt1Cre or Wnt1Cre2 driver to remove gene function throughout the neural crest, which gives rise to most of the facial bones (Danielian et al., 1998; Lewis et al., 2013). While extremely useful, these lines are not sufficient to address all questions relative to a gene’s function in the neural crest or in craniofacial development. Wnt1Cre/Wnt1Cre2 turn on after neural crest cells are specified (Barriga et al., 2015), and these lines are therefore unable to address issues relating to possible role of the gene under study in neural crest specification. Wnt1Cre/Wnt1Cre2 also label neural crest cells very broadly, and therefore are unable to address the roles of a specific gene in critical craniofacial subregions that can contribute to facial clefting phenotypes, such as those that we have observed with Pdgfra or Fgfr1 Wnt1Cre conditional mutants (Brewer et al., 2015; Tallquist and Soriano,
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影响因子：
64.5
作者：
Prescott SL;Srinivasan R;Marchetto MC;Grishina I;Narvaiza I;Selleri L;Gage FH;Swigut T;Wysocka J
通讯作者：
Wysocka J
影响因子：
10.5
作者：
Brewer JR;Molotkov A;Mazot P;Hoch RV;Soriano P
通讯作者：
Soriano P
影响因子：
30.8
作者：
Uslu, Veil Vural;Petretich, Massimo;Spitz, Francois
通讯作者：
Spitz, Francois
DOI：
10.1126/science.1241006
发表时间：
2013-10-25
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Attanasio C;Nord AS;Zhu Y;Blow MJ;Li Z;Liberton DK;Morrison H;Plajzer-Frick I;Holt A;Hosseini R;Phouanenavong S;Akiyama JA;Shoukry M;Afzal V;Rubin EM;FitzPatrick DR;Ren B;Hallgrímsson B;Pennacchio LA;Visel A
通讯作者：
Visel A
影响因子：
14.9
作者：
Hermann M;Stillhard P;Wildner H;Seruggia D;Kapp V;Sánchez-Iranzo H;Mercader N;Montoliu L;Zeilhofer HU;Pelczar P
通讯作者：
Pelczar P