Cis-regulatory elements such as enhancers play critical regulatory roles in modulating gene expression during brain development. The development of massively parallel reporter assays has enabled high-throughput functional screening of candidate DNA sequences for enhancer activity. Tissue-specific screening of in vivo enhancer function at scale has the potential to greatly expand our understanding of the role of non-coding sequences in development, evolution, and disease. Here, we adapted the self-transcribing regulatory element MPRA strategy for delivery to early postnatal mouse brain via recombinant adeno-associated virus (rAAV). We identify putative enhancers capable of driving reporter gene expression in mouse forebrain, including regulatory elements within an intronic CACNA1C linkage disequilibrium block associated with risk in neuropsychiatric disorder genetic studies. In vivo functional testing of putative enhancers, as we demonstrate here, will be critical toward characterizing regulatory activity of enhancers and understanding how enhancer sequences organize gene expression in normal and pathogenic brain development.

中文翻译：

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平行功能测试可鉴定出在出生后早期小鼠大脑中活跃的增强剂

顺式调节元件（例如增强子）在大脑发育过程中，在调节基因表达中起着关键的调节作用。大规模平行报告基因检测的发展已实现了候选DNA序列高通量功能筛选的增强子活性。大规模的体内增强子功能的组织特异性筛选有可能极大地扩展我们对非编码序列在发育，进化和疾病中的作用的理解。在这里，我们适应了通过转录腺相关病毒（rAAV）传递到产后早期小鼠大脑的自转录调控元件MPRA策略。我们确定了能够驱动小鼠前脑中的报告基因表达的推定增强子，包括与神经精神疾病遗传学研究中的风险相关的内含CACNA1C连锁不平衡区中的调控元件。正如我们在此处演示的那样，推定的增强子的体内功能测试对于表征增强子的调节活性以及了解增强子序列如何在正常和致病性大脑发育中组织基因表达至关重要。