MacKay Gillian E, Keighren Margaret A, Wilson Linda, Pratt Thomas, Flockhart Jean H, Mason John O, Price David J, West John D
Division of Reproductive and Developmental Sciences, University of Edinburgh, UK.
J Anat. 2005 Jan;206(1):79-92. doi: 10.1111/j.0021-8782.2005.00370.x.
The mouse TgTP6.3 transgene, encoding a tauGFP fusion protein, is becoming widely used but has yet to be fully characterized and evaluated as suitable lineage marker. The aim of the present study was to investigate the phenotype of TgTP6.3(+/+) homozygotes and TgTP6.3(+/-) hemizygotes, characterize the expression of the TgTP6.3 transgene in different tissues and critically evaluate its use as a lineage marker. TgTP6.3(+/+) homozygotes died between embryonic day 14.5 and weaning, whereas TgTP6.3(+/-) hemizygotes were mostly viable and fertile but smaller than non-transgenic siblings. TgTP6.3 expression began in the late two-cell stage, persisted in most fetal and adult tissues and was uniformly expressed in many (but not all) tissues. TgTP6.3(+/-) cells were readily identified in many chimeric tissues and their contribution appeared to be quantitatively and spatially normal. Overall, tauGFP expression in hemizygous TgTP6.3(+/-) cells fulfils the main criteria of a good lineage marker for many tissues. It provides a useful lineage marker, which should be particularly suitable for axons, blood vessels and pre-implantation embryos.
编码tauGFP融合蛋白的小鼠TgTP6.3转基因正被广泛使用,但作为一种合适的谱系标记物,其尚未得到充分表征和评估。本研究的目的是研究TgTP6.3(+/+)纯合子和TgTP6.3(+/-)半合子的表型,表征TgTP6.3转基因在不同组织中的表达,并严格评估其作为谱系标记物的用途。TgTP6.3(+/+)纯合子在胚胎第14.5天到断奶之间死亡,而TgTP6.3(+/-)半合子大多存活且可育,但比非转基因同胞小。TgTP6.3表达始于两细胞晚期,在大多数胎儿和成年组织中持续存在,并且在许多(但不是所有)组织中均匀表达。在许多嵌合组织中很容易鉴定出TgTP6.3(+/-)细胞,并且它们的贡献在数量和空间上似乎是正常的。总体而言,半合子TgTP6.3(+/-)细胞中的tauGFP表达满足了许多组织中良好谱系标记物的主要标准。它提供了一种有用的谱系标记物,特别适用于轴突、血管和植入前胚胎。