Single Cell Studies of Lineage Specific Expression of the Protocadherin Gene Clus

描述（由申请人提供）：成簇原钙粘蛋白（Pcdh）基因似乎是哺乳动物神经系统中单细胞多样性的主要来源。这个基因簇的复杂遗传组织表明，单细胞多样性可能是通过体细胞重组产生的，类似于在免疫球蛋白和T细胞受体中发现的，或者通过类似于在果蝇Dscam中发现的选择性剪接机制。令人惊讶的是，Pcdh同种型多样性似乎是通过随机启动子选择和替代前mRNA剪接的组合产生的。Pcdh多样性的这种模型主要基于来自一种神经元亚型的有限数量的数据。目前的建议的主要目的是开发和应用创新的单细胞表达谱和单分子原位检测方法，以检查该模型的有效性和/或一般性，研究Pcdh基因表达的复杂性在多种神经元细胞类型和发育阶段，并确定在神经发生Pcdh启动子选择发生的时间。

DESCRIPTION (provided by applicant): The clustered protocadherin (Pcdh) genes appear to be the primary source of single cell diversity in the mammalian nervous system. The complex genetic organization of this gene cluster suggested that single cell diversity might arise through somatic recombination, akin to that found in Immunoglobulin and T-Cell receptors or through an alternative splicing mechanism similar to that found in Dscam in Drosophila Melanogaster. Surprisingly, Pcdh isoform diversity appears to be generated through a combination of stochastic promoter choice and alternative pre-mRNA splicing. This model for Pcdh diversity is based primarily on a limited amount of data from one neuronal sub-type. The primary objective of the current proposal is to develop and apply innovative single cell expression profiling and single molecule in situ detection methods to examine the validity and/or generality of this model, to study the complexity of Pcdh gene expression in multiple neuronal cell types and developmental stages, and to determine the time during neurogenesis that Pcdh promoter choice occurs.