Department of Molecular and Cellular Biosciences, University of Cincinnati, Cincinnati, OH 45229, USA; Neuroscience Graduate Program, University of Cincinnati, Cincinnati, OH 45229, USA.
Department of Pediatrics, University of California, San Francisco, San Francisco, CA 94143, USA; Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94158, USA.
Cell Rep. 2024 Jan 23;43(1):113660. doi: 10.1016/j.celrep.2023.113660. Epub 2024 Jan 12.
The recent proliferation of new Cre and CreER recombinase lines provides researchers with a diverse toolkit to study microglial gene function. To determine how best to apply these lines in studies of microglial gene function, a thorough and detailed comparison of their properties is needed. Here, we examined four different microglial CreER lines (Cx3cr1, Cx3cr1, P2ry12, and Tmem119), focusing on (1) recombination specificity, (2) leakiness (the degree of tamoxifen-independent recombination in microglia and other cells), (3) the efficiency of tamoxifen-induced recombination, (4) extraneural recombination (the degree of recombination in cells outside of the CNS, particularly myelo/monocyte lineages), and (5) off-target effects in the context of neonatal brain development. We identify important caveats and strengths for these lines, which will provide broad significance for researchers interested in performing conditional gene deletion in microglia. We also provide data emphasizing the potential of these lines for injury models that result in the recruitment of splenic immune cells.
最近新的 Cre 和 CreER 重组酶系的大量出现为研究人员提供了一个多样化的工具包，用于研究小胶质细胞基因功能。为了确定如何最好地将这些线应用于小胶质细胞基因功能的研究，需要对它们的特性进行彻底和详细的比较。在这里，我们研究了四种不同的小胶质细胞 CreER 线（Cx3cr1、Cx3cr1、P2ry12 和 Tmem119），重点关注（1）重组特异性，（2）漏性（即他莫昔芬独立于小胶质细胞和其他细胞的重组程度），（3）他莫昔芬诱导的重组效率，（4）神经外重组（CNS 以外的细胞中的重组程度，特别是骨髓/单核细胞谱系），以及（5）在新生儿大脑发育背景下的脱靶效应。我们确定了这些线的重要注意事项和优势，这将为对在小胶质细胞中进行条件性基因缺失感兴趣的研究人员提供广泛的意义。我们还提供了强调这些线在导致脾脏免疫细胞募集的损伤模型中的潜力的数据。
