Efficient gene modulation in mouse epiblast using a Sox2Cre transgenic mouse strain
Efficient gene modulation in mouse epiblast using a Sox2Cre transgenic mouse strain
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DOI：
10.1016/s0925-4773(03)00099-6
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发表时间：
2002-12-01
影响因子：
2.6
通讯作者：
McMahon, Andrew P.
中科院分区：
文献类型：
--
作者：
Hayashi, Shigemi;Lewis, Paula;McMahon, Andrew P.
我们构建了一个转基因品系，该品系在来自Sox2基因的12.5kb上游调控序列的调控下表达Cre基因产物。利用R26R报告基因品系，我们表明该转基因品系在胚胎第6.5天（E6.5）可诱导所有上胚层细胞发生重组，但此时在其他胚外细胞类型中几乎没有或没有活性。当与音猬基因（Shh）的条件性等位基因（Shh(c)）杂交时，所有Sox2Cre；Shh(n)/Shh(c)胚胎表现出与Shh基因缺失突变体无法区分的表型。Sox2Cre在上胚层介导的重组中比Mox2Cre（MORE）转基因品系更有效，Mox2Cre转基因品系也已被证明仅在早期小鼠胚胎的胚胎部分驱动Cre介导的重组。尽管大多数MORE；shh(h)/shh(c)胚胎具有shh缺失表型，但33%表现出较温和的骨骼表型，这很可能是卵柱阶段重组不完全的结果。与这些发现一致的是，在原肠胚早期阶段，Sox2Cre更早发挥作用，且Sox2Cre介导的重组比MORE介导的重组更先进。Sox2Cre品系可能在实现基因活性的完全、上胚层特异性去除方面更有效，而MORE品系的嵌合活性将有助于在胚胎本体中产生部分功能丧失表型。（C）2003 Elsevier Science Ireland Ltd.保留所有权利。
We have generated a transgenic line that expresses the Cre gene product under the regulation of a 12.5 kb upstream regulatory sequence from the Sox2 gene. Using a R26R reporter line, we show that this transgenic line induces recombination in all epiblast cells by embryonic day ( E) 6.5 but little or no activity in other extraembryonic cell types at this time. When crossed to a conditional allele of the Sonic hedgehog gene ( Shh(c)), all Sox2Cre; Shh(n)/Shh(c) embryos displayed a phenotype indistinguishable from that of the Shh null mutant. Sox2Cre functioned more efficiently in epiblast-mediated recombination than the Mox2Cre ( MORE) transgenic line, which has also been shown to drive Cre-mediated recombination exclusively in the embryonic component of the early mouse embryo. Although most MORE; shh(h)/shh(c) embryos have a shh hull phenotype, 33% displayed a milder skeletal phenotype, most likely result of incomplete recombination at egg cylinder stages. In agreement with these findings, Sox2Cre was active earlier and Sox2Cre-mediated recombination was more advanced than MORE-mediated recombination at early gastrulation stages. The Sox2Cre line is likely to be more effective in generating complete, epiblast-specific removal of gene activity, and the mosaic activity of the MORE line will be helpful in generating partial loss-of-function phenotypes in the embryo-proper. (C) 2003 Elsevier Science Ireland Ltd. All rights reserved.