State Key Laboratory of Animal Nutrition, Key Laboratory of Farm Animal Genetic Resource and Germplasm Innovation of Ministry of Agriculture, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R. China.
Animal Functional Genomics Group, Agricultural Genomes Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, Guangdong 518120, P.R. China.
Mol Med Rep. 2018 Jan;17(1):37-50. doi: 10.3892/mmr.2017.7842. Epub 2017 Oct 20.
Multi‑transgenic technology is superior to single transgenic technology in biological and medical research. Multi‑transgene insertion mediated by a polycistronic system is more effective for the integration of polygenes. The multi‑transgene insertion patterns and manners of inheritance are not completely understood. Copy number quantification is one available approach for addressing this issue. The present study determined copy numbers in two multi‑transgenic mice (K3 and L3) and two multi‑transgenic miniature pigs (Z2 and Z3) using absolute quantitative polymerase chain reaction analysis. For the F0 generation, a given transgene was able to exhibit different copy number integration capacities in different individuals. For the F1 generation, the most notable characteristic was that the copy number proportions were different among pedigrees (P<0.05). The results of the present study demonstrated that transgenes within the same vector exhibited the same integration trend between the F0 and F1 generations. In conclusion, intraspecific consistency and intergenerational copy numbers were compared and the integration capacity of each specific transgene differed in multi‑transgenic animals. In particular, the copy number of one transgene may not be used to represent other transgenes in polycistronic vector‑mediated multi‑transgenic organisms. Consequently, in multi‑transgenic experimental animal disease model research or breeding, copy numbers provide an important reference. Therefore, each transgene in multi‑transgenic animals must be separately screened to prevent large copy number differences, and inconsistent expression between transgenes and miscellaneous data, in subsequent research.
多转基因技术在生物和医学研究中优于单转基因技术。多顺反子系统介导的多转基因插入对于多基因的整合更为有效。多转基因的插入模式和遗传方式尚不完全清楚。拷贝数定量是解决这一问题的一种可行方法。本研究采用绝对定量聚合酶链反应分析，测定了两只多转基因小鼠（K3 和 L3）和两只多转基因小型猪（Z2 和 Z3）的拷贝数。对于 F0 代，一个给定的转基因在不同个体中能够表现出不同的拷贝数整合能力。对于 F1 代，最显著的特征是不同家系之间的拷贝数比例不同（P<0.05）。本研究结果表明，同一载体中的转基因在 F0 和 F1 代之间表现出相同的整合趋势。总之，进行了种内一致性和代际拷贝数的比较，多转基因动物中每个特定转基因的整合能力不同。特别是，一个转基因的拷贝数不能用于代表多顺反子载体介导的多转基因生物中的其他转基因。因此，在多转基因实验动物疾病模型研究或繁殖中，拷贝数提供了重要的参考。因此，在多转基因动物中，每个转基因都必须单独筛选，以防止在后续研究中出现大的拷贝数差异、转基因之间的不一致表达和杂散数据。
