Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice.
Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice.
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DOI：
10.1242/bio.058678
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发表时间：
2021-09-15
期刊：
影响因子：
2.4
通讯作者：
Bird TG
中科院分区：
文献类型：
--
作者：
Kiourtis C;Wilczynska A;Nixon C;Clark W;May S;Bird TG
小鼠是一种广泛使用的临床前模型系统，这在很大程度上是由于其遗传操作的潜力。在时间控制下操纵特定细胞中基因表达的能力是一种强大的实验工具。肝脏是代谢稳态的中心，也是许多疾病的发病部位，这使得肝细胞的靶向具有吸引力。腺相关病毒8（AAV 8）载体是操纵肝细胞基因表达的有价值的工具。然而，它们在小鼠中的脱靶效应尚未得到彻底研究。在此，我们试图鉴定AAV 8施用在小鼠中的短期脱靶效应。为了做到这一点，我们用表达Cre重组酶的重组AAV 8载体或对照AAV 8载体注射C57 BL/6 J野生型小鼠，并表征与未注射对照相比一般健康和肝脏生理学、组织学和转录组学的变化。我们观察到AAV 8给药后稳态肝脏增殖减少以及DNA损伤标志物γ H2 AX诱导的急性和短暂趋势。后者在Cre重组酶表达后通过载体增强。此外，我们还观察到与昼夜节律和感染反应有关的基因的转录变化。值得注意的是，在通过AAV 8载体表达Cre重组酶时没有额外的转录组学变化。总体而言，没有肝损伤的证据，并且在AAV 8感染后14天仅观察到轻度T细胞浸润。 这些数据推进了使用Cre表达AAV载体通过Cre-Lox重组进行肝细胞基因组编辑的技术，证明了它们对鼠生理学的最小影响，并突出了这些系统的更微妙的脱靶效应。总结：本文提供了一个全面的特点，对小鼠生理学，肝组织学和肝转录组的短期影响的腺相关病毒8管理。
Mice are a widely used pre-clinical model system in large part due to their potential for genetic manipulation. The ability to manipulate gene expression in specific cells under temporal control is a powerful experimental tool. The liver is central to metabolic homeostasis and a site of many diseases, making the targeting of hepatocytes attractive. Adeno-associated virus 8 (AAV8) vectors are valuable instruments for the manipulation of hepatocellular gene expression. However, their off-target effects in mice have not been thoroughly explored. Here, we sought to identify the short-term off-target effects of AAV8 administration in mice. To do this, we injected C57BL/6J wild-type mice with either recombinant AAV8 vectors expressing Cre recombinase or control AAV8 vectors and characterised the changes in general health and in liver physiology, histology and transcriptomics compared to uninjected controls. We observed an acute and transient trend for reduction in homeostatic liver proliferation together with induction of the DNA damage marker γH2AX following AAV8 administration. The latter was enhanced upon Cre recombinase expression by the vector. Furthermore, we observed transcriptional changes in genes involved in circadian rhythm and response to infection. Notably, there were no additional transcriptomic changes upon expression of Cre recombinase by the AAV8 vector. Overall, there was no evidence of liver injury, and only mild T-cell infiltration was observed 14 days following AAV8 infection. These data advance the technique of hepatocellular genome editing through Cre-Lox recombination using Cre expressing AAV vectors, demonstrating their minimal effects on murine physiology and highlight the more subtle off target effects of these systems. Summary: This paper provides a comprehensive characterisation of the short-term effects of administration of adeno-associated virus 8 on murine physiology, liver histology and liver transcriptome.
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影响因子：
32.4
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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DOI：
10.1073/pnas.2004838117
发表时间：
2021-06-01
影响因子：
11.1
作者：
Garrood, William T.;Kranjc, Nace;Simoni, Alekos
通讯作者：
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