Transgenic mice expressing Cre-recombinase specifically in retinal rod bipolar neurons.
Transgenic mice expressing Cre-recombinase specifically in retinal rod bipolar neurons.
复制标题
转基因小鼠在视网膜杆状双极神经元中特异性表达 Cre 重组酶。
DOI：
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发表时间：
2005
影响因子：
4.4
通讯作者：
Jian
中科院分区：
文献类型：
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作者：
Xin;Baiyu Chen;Alam Hoi;J. Tanner;D. Tay;K. So;R. Rachel;N. Copeland;N. Jenkins;Jian
目的
建立在视网膜视杆双极细胞中表达Cre重组酶的转基因小鼠品系，用于产生视杆双极细胞特异性敲除突变体。
方法
通过red介导的重组工程，将IRES-Cre-cDNA片段插入到携带完整Pcp2基因的173-kb细菌人工染色体（BAC）中。用修饰的BAC产生转基因小鼠并鉴定。将Cre转基因小鼠与ROSA 26和Z/EG报告小鼠杂交以检测Cre重组酶活性。
结果
X-gal染色显示，视网膜内核层和小脑浦肯野细胞中有较强的Cre重组酶活性。用抗GFP抗体和抗PKCalpha抗体（对视网膜视杆双极细胞特异性）的双重染色揭示Cre重组酶活性仅定位于视网膜中的视杆双极细胞。
结论
建立了在视网膜杆双极神经元中表达Cre重组酶的小鼠BAC-Pcp2-IRES-Cre转基因系。由于一些广泛表达基因的突变可能导致视网膜变性疾病，小鼠品系BAC-Pcp2-IRES-Cre将是一个有用的新工具，研究视网膜杆双极细胞特异性基因失活的影响。
PURPOSE
To establish a transgenic mouse line that expresses Cre-recombinase in retinal rod bipolar cells for the generation of rod bipolar cell-specific knockout mutants.
METHODS
The IRES-Cre-cDNA fragment was inserted into a 173-kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene, by using red-mediated recombineering. Transgenic mice were generated with the modified BAC and identified. The Cre-transgenic mice were crossed with ROSA26 and Z/EG reporter mice to detect Cre-recombinase activity.
RESULTS
X-gal staining showed that strong Cre-recombinase activities were present in retinal inner nuclear layers and cerebellar Purkinje cells. Double staining with an anti-GFP antibody and an anti-PKCalpha antibody (specific for retinal rod bipolar cells) revealed that Cre-recombinase activity localized exclusively to the rod bipolar cells in the retina.
CONCLUSIONS
A mouse BAC-Pcp2-IRES-Cre transgenic line that expresses Cre-recombinase in retinal rod bipolar neurons has been established. Because mutations in some ubiquitously expressed genes may result in retinal degenerative diseases, the mouse strain BAC-Pcp2-IRES-Cre will be a useful new tool for investigating the effects of retinal rod bipolar cell-specific gene inactivation.