A knock-in allele of En1 expressing dre recombinase.
A knock-in allele of En1 expressing dre recombinase.
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DOI：
10.1002/dvg.22954
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发表时间：
2016-08
期刊：
影响因子：
--
通讯作者：
Jensen P
中科院分区：
文献类型：
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作者：
Plummer NW;de Marchena J;Jensen P
EN1是一种含有同源盒的转录因子，在发育过程中在不同的组织中表达，包括胚胎中脑和后脑的前部。为了便于研究具有EN1表达历史的细胞之间的遗传和发育异质性，我们产生了En1Dre，一种表达DRE重组酶的敲入等位基因。En1Dre可以与现有的Cre和FLP重组酶系一起用于遗传交叉标记、命运图谱和以瞬时表达EN1为特征的细胞亚群的功能操纵。为了避免干扰EN1的功能，在EN1编码序列的3‘端插入了DRE cDNA，与病毒2A肽一起介导了单独的EN1和DRE蛋白的翻译。因此，活的和可育的En1Dre纯合子可以用来增加在复杂杂交中产生的有用基因型的比例。在妊娠中期的小鼠胚胎中，En1Dre的Dre表达模式与野生型EN1的表达模式没有区别，En1Dre通过在体内有效地重组Rox位点来控制Dre响应的指示等位基因。通过应用基因工具，允许基于多种不同重组酶的组合表达来操纵细胞，En1Dre将显著扩展针对更广泛的EN1表达区域内的重要神经元亚群和其他细胞的能力。
En1 is a homeobox-containing transcription factor expressed during development in diverse tissues, including the embryonic midbrain and anterior hindbrain. To facilitate investigation of genetic and developmental heterogeneity among cells with a history of En1 expression, we have generated En1Dre, a knock-in allele expressing Dre recombinase. En1Dre can be used with existing Cre and Flp recombinase lines for genetic intersectional labeling, fate mapping, and functional manipulation of subpopulations of cells characterized by transient expression of En1. To avoid disrupting En1 function, the Dre cDNA is inserted at the 3′ end of the En1 coding sequence, together with a viral 2A peptide to mediate translation of separate EN1 and Dre proteins. Consequently, viable and fertile En1Dre homozygotes can be used to increase the proportion of useful genotypes produced in complex crosses. The pattern of Dre expression from En1Dre is indistinguishable from wild-type En1 expression in mid-gestation mouse embryos, and En1Dre controls Dre-responsive indicator alleles by efficiently recombining rox sites in vivo. Through the application of genetic tools that allow manipulation of cells based on combinatorial expression of multiple distinct recombinases, En1Dre will significantly extend the ability to target important subpopulations of neurons and other cells within the broader En1 expression domain.