Genomic Analysis of Nucleic Acid Transactions

核酸交易的基因组分析

• 批准号: 10596116
• 负责人: Brenton R. Graveley
• 金额: $ 63.24万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10596116
• 关键词: Adaptive Immune System; Address; Alternative Splicing; Basic Science; CRISPR screen; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; DNA Sequence; Development; Elements; Future; Gene Expression; Gene Proteins; Genes; Genetic Transcription; Genome; Genomics; Guide RNA; Health; Human; Immune system; Immunologic Memory; Invaded; Messenger RNA; Nucleic Acids; Organism; Plasmids; Process; Prokaryotic Cells; Proteins; Proteome; RNA Processing; RNA Splicing; RNA interference screen; RNA-Binding Proteins; Reporter; Role; System; Virus; Work; experimental study; genome editing; innovation; insight; mRNA Precursor; memory encoding; nanopore; nuclease; single-cell RNA sequencing; tool

Most eukaryotic pre-mRNAs, especially in metazoans, are alternatively spliced to generate multiple mRNAs and proteins. Given the importance of alternative splicing in regulating gene expression and enhancing the diversity of the proteome, it is essential to understand the mechanisms of splicing and how alternative splicing is regulated. In this project, we will study the roles of RNA binding proteins in alternative splicing, with an emphasis on how RNA binding proteins auto- and cross-regulate the splicing their own and other RNA binding protein genes. This work will provide new insight into the mechanisms of RNA processing and how these proteins regulate one another to achieve homoestasis. Many prokaryotes encode CRISPR-Cas systems which are RNA-guided adaptive immune systems that protects prokaryotic organisms against invaders such as viruses and plasmids. Immune memories are encoded as short DNA sequences, called “spacers”, that match invader genomes and are stored as interspersed elements in an array of short repeats (the CRISPR array). The CRISPR arrays are transcribed and processed into guide RNAs which pair with Cas nucleases to recognize and degrade target nucleic acid (interference). New immune memories are formed during “adaptation” when fragments of invader DNA are acquired and integrated into CRISPR arrays for use in future targeting. While a tremendous amount is known about the targeting and degradation of invading nucleic acids, much less is known about the process of adaptation. We plan to further characterize the adaptation process in prokaryotic CRISPR-Cas systems. This work will also provide insight into the mechanisms of adaptation in the immune systems of prokaryotes. In addition to enhancing our understanding of the basic science of prokaryotic immune systems, there is tremendous potential that this work could lead to the development of new tools that can be used for genome editing applications. All of these projects will be addressed using the types of general approaches we have developed such as splicing reporters, single cell RNA-Seq, nanopore sequencing, RNAi or CRISPR screens, and computational genomics. We will also continue to develop additional innovative approaches to address these issues as needed or as opportunities arise due to technical advances in the field.

大多数真核生物的前mRNA，尤其是在后生动物中，是选择性剪接以产生多个 mRNA和蛋白质。鉴于选择性剪接在调节基因表达和增强 由于蛋白质组的多样性，了解剪接的机制以及选择性剪接是如何发生的是至关重要的。 剪接受到调节。在这个项目中，我们将研究RNA结合蛋白在选择性剪接中的作用， 强调RNA结合蛋白如何自动和交叉调节其自身和其他RNA的剪接 结合蛋白基因这项工作将提供新的见解RNA加工的机制，以及如何 这些蛋白质相互调节以实现体内稳态。 许多原核生物编码CRISPR-Cas系统，其是RNA引导的适应性免疫系统， 保护原核生物免受病毒和质粒等入侵者的侵害。免疫记忆是 编码为短的DNA序列，称为“间隔区”，与入侵者基因组相匹配，并储存为 短重复序列阵列（CRISPR阵列）中的散布元件。CRISPR阵列被转录 并加工成与Cas核酸酶配对以识别和降解靶核酸的指导RNA （干扰）。新的免疫记忆是在“适应”过程中形成的， 获得并整合到CRISPR阵列中用于未来的靶向。虽然我们知道 关于入侵核酸的靶向和降解，对入侵核酸的过程知之甚少。 适应我们计划进一步表征原核CRISPR-Cas系统中的适应过程。这 这项工作还将使人们深入了解原核生物免疫系统的适应机制。在 除了增强我们对原核免疫系统基础科学的理解外， 巨大的潜力，这项工作可以导致新的工具，可用于基因组的发展 编辑应用程序。 所有这些项目都将使用我们开发的一般方法类型进行处理， 作为剪接报告基因、单细胞RNA-Seq、纳米孔测序、RNAi或CRISPR筛选，以及 计算基因组学我们还将继续开发其他创新方法来解决这些问题。 根据需要或随着该领域的技术进步而出现的机会来解决问题。