Institute for Human Genetics, University of California, San Francisco, CA, USA.
Department of Neurology, University of California, San Francisco, CA, USA.
Hum Mol Genet. 2022 Oct 20;31(R1):R47-R53. doi: 10.1093/hmg/ddac193.
Advances in genomics have led to the identification of many risk loci with hundreds of genes and thousands of DNA variants associated with neuropsychiatric disorders. A significant barrier to understanding the genetic underpinnings of complex diseases is the lack of functional characterization of risk genes and variants in biological systems relevant to human health and connecting disease-associated variants to pathological phenotypes. Characterizing gene and DNA variant functions requires genetic perturbations followed by molecular and cellular assays of neurobiological phenotypes. However, generating null or mutant alleles is low throughput, making it impossible to characterize disease-associated variants in large quantities efficiently. CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) screens can be leveraged to dissect the biological consequences of the tested genes and variants in their native context. Nevertheless, testing non-coding variants associated with complex diseases remains non-trivial. In this review, we first discuss the current challenges of interpreting the function of the non-coding genome and approaches to prioritizing disease-associated variants in the context of the 3D epigenome. Second, we provide a brief overview of high-throughput CRISPRi and CRISPRa screening strategies applicable for characterizing non-coding sequences in appropriate biological systems. Lastly, we discuss the promising prospects of using CRISPR-based technologies to dissect DNA sequences associated with neuropsychiatric diseases.
基因组学的进步导致了许多风险基因座的鉴定，这些基因座与数百个基因和数千个与神经精神疾病相关的 DNA 变体有关。理解复杂疾病遗传基础的一个显著障碍是缺乏对与人类健康相关的生物系统中风险基因和变体的功能表征，以及将疾病相关变体与病理表型联系起来。表征基因和 DNA 变体的功能需要进行遗传扰动，然后对神经生物学表型进行分子和细胞分析。然而，产生缺失或突变等位基因的效率很低，使得无法有效地对大量疾病相关变体进行特征描述。CRISPR 干扰（CRISPRi）和 CRISPR 激活（CRISPRa）筛选可以用于剖析在其天然环境中测试的基因和变体的生物学后果。然而，测试与复杂疾病相关的非编码变体仍然具有挑战性。在这篇综述中，我们首先讨论了目前解释非编码基因组功能的挑战，以及在 3D 表观基因组背景下优先考虑疾病相关变体的方法。其次，我们简要概述了适用于在适当的生物系统中对非编码序列进行高通量 CRISPRi 和 CRISPRa 筛选的策略。最后，我们讨论了使用基于 CRISPR 的技术剖析与神经精神疾病相关的 DNA 序列的有前途的前景。
