Nobody Is Perfect: Cre Drivers Deserve Careful Consideration.
Nobody Is Perfect: Cre Drivers Deserve Careful Consideration.
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没有人是完美的：Cre 驱动程序值得仔细考虑。
DOI：
10.1161/atvbaha.123.319683
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Gomez,Delphine
中科院分区：
文献类型：
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作者：
Gomez,Delphine
用于细胞特异性和时间控制重组的 Cre 驱动程序的开发已经改变了急性和慢性血管疾病背景下血管平滑肌细胞 (SMC) 研究的游戏规则。雌激素受体融合的 Cre 重组酶的产生，实现了他莫昔芬依赖性核易位和 floxed 位点切除，避免了胚胎和出生后基因敲除致死率，同时允许在血管疾病发病机制和成体生物体细胞命运图谱期间对基因功能进行更严格的评估。 1 细胞特异性和时间控制的 Cre 驱动程序的效率、特异性和可靠性取决于 (1) 用于驱动 Cre 表达的启动子的细胞特异性，(2) 启动子活性和 Cre 表达水平，以及 (3) loxP 位点切除需要他莫昔芬给药的程度，包括相关的缺乏他莫昔芬独立重组。考虑到这些因素，Offermanns 实验室培育出了 Myh11-CreERT2 转基因小鼠，该小鼠很快成为可诱导 SMC 特异性敲除和命运图谱的黄金标准，并且在过去十年中一直如此。 2 Myh11-CreERT2 转基因包含在细菌人工染色体中，该染色体包含小鼠 Myh11 基因体，并在 Myh11 起始密码子处插入了 CreERT2 盒。尽管最初的报告中没有提及，但后来注意到 Myh11-CreERT2 转基因具有 Y 连锁遗传模式，表明细菌人工染色体随机整合在 Y 染色体内。 3 使用基于 Myh11-CreERT2 的基因删除和命运图谱进行范式转变观察，导致重新评估 SMC 在血管疾病中的参与和功能。值得注意的是，这些研究证明了 SMC 在动脉粥样硬化中的广泛可塑性及其向多种不同表型状态的依赖环境的转变。 4
The development of Cre drivers for cell-specific and temporally controlled recombination has been a game changer for studying vascular smooth muscle cells (SMCs) in the context of acute and chronic vascular diseases. The generation of the estrogen receptor–fused Cre recombinase, enabling tamoxifen-dependent nuclear translocation and floxed site excisions, circumvented embryonic and postnatal knockout lethality while allowing for more rigorous assessment of gene function during vascular disease pathogenesis and cell fate mapping in adult organisms. 1 The efficiency, specificity, and reliability of cell-specific and temporally controlled Cre drivers depend on (1) the cell specificity of the promoter used to drive Cre expression,(2) the promoter activity and Cre expression levels, and (3) the extent to which loxP site excision requires tamoxifen administration, including an associated lack of tamoxifen-independent recombination. Considering these factors, the Offermanns laboratory generated the Myh11-CreERT2 transgenic mouse, which quickly became the gold standard for inducible SMC-specific knockout and fate mapping and has remained so for the past decade. 2 The Myh11-CreERT2 transgene is contained in a bacterial artificial chromosome comprising the mouse Myh11 gene body with the CreERT2 cassette inserted at the Myh11 start codon. Although not mentioned in the initial report, the Myh11-CreERT2 transgene was later noted to have a Y-linked inheritance pattern, suggesting random integration of the bacterial artificial chromosome within the Y chromosome. 3 Paradigm-shifting observations have been made using Myh11-CreERT2–based gene deletion and fate mapping, leading to a reevaluation of the participation and functions of SMC in vascular disease. Notably, these studies demonstrated extensive plasticity of SMCs in atherosclerosis and their context-dependent transitions to multiple distinct phenotypic states. 4