Single Cell Genomic DNA Analysis in Central Nervous System (CNS)

Introduction of Single Cell Genomic DNA Analysis

Single cell genomics refers to the study of genetic heterogeneity induced from the stochastic intercellular variation of the genome, proteome, metabolome, and epigenome. Based on the development of advanced technologies in genomics, biochemistry, and molecular biology, single cell analysis has opened new frontiers for large-scale studies of genome organization and behavior. In recent years, some studies showed that single cell genomic DNA analysis presents great potentials in the research of human neuronal diversity and brain diseases. Human neuronal diversity is thought to be mediated by somatic mutations in the genome and epigenome. In the central nervous system (CNS), intercellular epigenome variations mediated by changes of gene expression profiles appear to be related to a series of neurodevelopmental, neuropsychiatric, and neurodegenerative disorders.

Techniques for Single Cell Genomic Analysis in CNS

For single cell genomic DNA analysis in CNS, there are a series of techniques that can be divided into two main categories: macromolecules analysis methods through cell isolation and direct macromolecules imaging methods in individual cells. Like other cell types or tissues, the isolation of macromolecular fractions from brain tissues is required for the analysis of genetic and biochemical processes. It is worth mentioning that the novel lab-on-chip technology has been served as a powerful tool for single cell genomic analysis.

• direct staining of cells or subcellular structures
• DNA/RNA interphase fluorescence in situ hybridization (FISH)
• immunohistochemical analysis
• immunocytochemical analysis
• array comparative genomic hybridization (CGH)
• qPCR/RT-qPCR
• capillary electrophoresis
• transcriptome and proteome microarrays
• genomic sequencing
• mass spectrometry- and nuclear magnetic resonance-based techniques

Single Cell Genomic of Neurological Diseases

It is reported that somatic genome mutations are related to non-malignant brain diseases through different mechanisms of action. Using the single cell genomics approach, a series of molecular and cellular pathways of neurological and psychiatric diseases have been observed. For example, the brain-specific aneuploidy of chromosome 21 is an element in the Alzheimer’s disease (AD) neurodegeneration cascade. 410 unique copy number variations (CNVs) can be found in the patients with Amyotrophic Lateral Sclerosis (Sporadic). There is low-level chromosomal mosaicism in 16% of children with idiopathic autism. Tissue-specific trinucleotide repeat (CAG) expansions in specific neuronal populations are associated with Huntington's disease. Rett Syndrome is induced by somatic mosaicism for MEPC2 mutations in females.

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