Simple and rapid determination of homozygous transgenic mice via in vivo fluorescence imaging.
Simple and rapid determination of homozygous transgenic mice via in vivo fluorescence imaging.
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通过体内荧光成像简单快速地确定纯合转基因小鼠
DOI：
10.18632/oncotarget.5535
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发表时间：
2015-11-17
期刊：
影响因子：
--
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Lin X;Jia J;Qin Y;Lin X;Li W;Xiao G;Li Y;Xie R;Huang H;Zhong L;Wu Q;Wang W;Huang W;Yao K;Xiao D;Sun Y
制定转基因小鼠品系的育种计划需要区分纯合子和杂合子转基因动物。将荧光报告转基因技术与小动物体内成像系统相结合，可以快速、直观地对转基因纯合子小鼠(S)进行体内荧光成像。RLG、RCLG或Rm17LG转基因小鼠普遍表达红色荧光蛋白(RFP)。为了鉴定纯合子的RLG转基因小鼠，对来自同一转基因创始人的RFP阳性杂合转基因小鼠(F1代)交配产生的所有新生F2代小鼠进行了全身荧光成像。随后，进行了体内荧光成像的即时数据分析，极大地方便了我们从F2代产仔中快速区分出具有强荧光的转RLG基因个体(S)，然后进一步确定这个/这些具有强荧光的RLG个体(S)为纯合子，这一点经小鼠交配得到了强烈的证实。此外，上述光学方法还可以快速、准确地区分纯合子RCLG或Rm17LG转基因小鼠。通过小鼠交配，在最短的时间内分别获得了10只、8只和2只RLG、RCLG和Rm17LG转基因纯合子小鼠，表明了这种光学方法的实用性和可靠性。综上所述，我们的研究结果充分表明，在本研究所述的情况下，体内荧光成像为在普遍存在的启动子控制下鉴定携带荧光报告基因的纯合子转基因小鼠提供了一种直观、快速和可靠的替代传统方法(即小鼠交配和实时定量PCR)。
Setting up breeding programs for transgenic mouse strains require to distinguish homozygous from the heterozygous transgenic animals. The combinational use of the fluorescence reporter transgene and small animal in-vivo imaging system might allow us to rapidly and visually determine the transgenic mice homozygous for transgene(s) by the in vivo fluorescence imaging. RLG, RCLG or Rm17LG transgenic mice ubiquitously express red fluorescent protein (RFP). To identify homozygous RLG transgenic mice, whole-body fluorescence imaging for all of newborn F2-generation littermates produced by mating of RFP-positive heterozygous transgenic mice (F1-generation) derived from the same transgenic founder was performed. Subsequently, the immediate data analysis of the in vivo fluorescence imaging was carried out, which greatly facilitated us to rapidly and readily distinguish RLG transgenic individual(s) with strong fluorescence from the rest of F2-generation littermates, followed by further determining this/these RLG individual(s) showing strong fluorescence to be homozygous, as strongly confirmed by mouse mating. Additionally, homozygous RCLG or Rm17LG transgenic mice were also rapidly and precisely distinguished by the above-mentioned optical approach. This approach allowed us within the shortest time period to obtain 10, 8 and 2 transgenic mice homozygous for RLG, RCLG and Rm17LG transgene, respectively, as verified by mouse mating, indicating the practicality and reliability of this optical method. Taken together, our findings fully demonstrate that the in vivo fluorescence imaging offers a visual, rapid and reliable alternative method to the traditional approaches (i.e., mouse mating and real-time quantitative PCR) in identifying homozygous transgenic mice harboring fluorescence reporter transgene under the control of a ubiquitous promoter in the situation mentioned in this study.