An AAV-CRISPR/Cas9 strategy for gene editing across divergent rodent species: Targeting neural oxytocin receptors as a proof of concept

一种跨不同啮齿动物物种进行基因编辑的 AAV-CRISPR/Cas9 策略:以神经催产素受体为目标作为概念验证

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作者:Arjen J Boender, Marina Boon, H Elliott Albers, Samantha R Eck, Brandon A Fricker, Aubrey M Kelly, Joseph E LeDoux, Simone C Motta, Prerana Shrestha, Jack H Taylor, Brian C Trainor, Rodrigo Triana-Del Rio, Larry J Young

Abstract

A major issue in neuroscience is the poor translatability of research results from preclinical studies in animals to clinical outcomes. Comparative neuroscience can overcome this barrier by studying multiple species to differentiate between species-specific and general mechanisms of neural circuit functioning. Targeted manipulation of neural circuits often depends on genetic dissection, and use of this technique has been restricted to only a few model species, limiting its application in comparative research. However, ongoing advances in genomics make genetic dissection attainable in a growing number of species. To demonstrate the potential of comparative gene editing approaches, we developed a viral-mediated CRISPR/Cas9 strategy that is predicted to target the oxytocin receptor (Oxtr) gene in >80 rodent species. This strategy specifically reduced OXTR levels in all evaluated species (n = 6) without causing gross neuronal toxicity. Thus, we show that CRISPR/Cas9-based tools can function in multiple species simultaneously. Thereby, we hope to encourage comparative gene editing and improve the translatability of neuroscientific research.

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