Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294.
Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Birmingham, AL 35294
J Immunol. 2020 Apr 1;204(7):1982-1987. doi: 10.4049/jimmunol.1901338. Epub 2020 Mar 2.
GFP is frequently used as a marker for tracking donor cells adoptively transplanted into recipient animals. The human ubiquitin C promoter (UBC)-driven-GFP transgenic mouse is a commonly used source of donor cells for this purpose. This mouse was initially generated in the C57BL/6 inbred strain and has been backcrossed into the BALB/cBy strain for over 11 generations. Both the C57BL/6 inbred and BALB/cBy congenic UBC-GFP lines are commercially available and have been widely distributed. These UBC-GFP lines can be a convenient resource for tracking donor cells in both syngenic MHC-matched and in allogenic MHC-mismatched studies as C57BL/6 (H-2) and BALB/cBy (H-2) have disparate MHC haplotypes. In this report, we surprisingly discover that the UBC-GFP BALB/cBy congenic mice still retain the H-2 MHC haplotype of their original C57BL/6 founder, suggesting that the UBC-GFP transgene integration site is closely linked to the MHC locus on chromosome 17. Using linear amplification-mediated PCR, we successfully map the UBC-GFP transgene to the MHC locus. This study highlights the importance and urgency of mapping the transgene integration site of transgenic mouse strains used in biomedical research. Furthermore, this study raises the possibility of alternative interpretations of previous studies using congenic UBC-GFP mice and focuses attention on the necessity for rigor and reproducibility in scientific research.
GFP 常被用作示踪剂,用于追踪被主动移植到受者动物体内的供体细胞。人泛素 C 启动子(UBC)驱动 GFP 转基因小鼠是用于此目的的常用供体细胞来源。这种小鼠最初是在 C57BL/6 近交系中产生的,并已回交至 BALB/cBy 近交系超过 11 代。C57BL/6 近交系和 BALB/cBy 同基因 UBC-GFP 系均可商业购买,并已广泛分布。这些 UBC-GFP 系可作为追踪同种 MHC 匹配和异种 MHC 不匹配研究中供体细胞的便利资源,因为 C57BL/6(H-2)和 BALB/cBy(H-2)具有不同的 MHC 单倍型。在本报告中,我们惊讶地发现,UBC-GFP BALB/cBy 同基因小鼠仍然保留其原始 C57BL/6 创始人的 H-2 MHC 单倍型,这表明 UBC-GFP 转基因的整合位点与 17 号染色体上的 MHC 基因座紧密连锁。使用线性扩增介导 PCR,我们成功地将 UBC-GFP 转基因定位到 MHC 基因座。本研究强调了在生物医学研究中对用于转基因小鼠系的转基因整合位点进行作图的重要性和紧迫性。此外,本研究提出了使用同基因 UBC-GFP 小鼠进行先前研究的替代解释的可能性,并引起了对科学研究中严谨性和可重复性的关注。