High throughput RNAi based functional genomics in primary cells and in vivo

原代细胞和体内基于高通量 RNAi 的功能基因组学

基本信息

批准号：
8311411
负责人：
Alexey Wolfson
金额：
$ 28.14万
依托单位：
ADVANCED RNA TECHNOLOGIES, LLC
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2014-05-01
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8311411
关键词：
Algorithms Area Automation Base Sequence Bioinformatics Biomedical Research Cells Chemistry Collaborations Communities Complex Custom Data Set Databases Development Drug Formulations Elements Environment Equipment and supply inventories Funding Gene Silencing Genes Genetic Transcription Genomics Goals Human Genome Immunology In Vitro Inflammation Inflammatory Libraries Maps Messenger RNA Methods Neurobiology Organ Model Pathway interactions Pharmacologic Substance Phase Process RNA RNA Interference Reagent Reporter Research Research Infrastructure Research Methodology Research Personnel Screening procedure Small Interfering RNA Stem cells Structure System Technology Time Tissue Model Tissues Transcript Transcriptional Regulation Transfection Validation base blastomere structure cell type commercialization cost design established cell line functional genomics gene function high throughput screening improved in vivo in vivo Model novel oncology parallel processing programs scaffold success text searching tool uptake web based interface

项目摘要

DESCRIPTION (provided by applicant): Completion of the HGP (human genome program) has opened the 'Genomic Era' in biomedical research and enables functional genomics analysis of various gene elements. The long-term goal of the ENCODE Project is to identify and functionally characterize all of the sequence-based genomic elements. Functional elements that have been studied in ENCODE include transcribed sequences, regulators of transcription, and regulators of RNA transcripts themselves. The function of more than 50% of transcribed gene sequences is unknown. Most of them are expressed in specialized cells and thus their functions can only be studied in a biologically relevant context (i.e. primary cells, in vivo). In this propoal we plan to develop and commercialize a new RNAi platform that enables gene studies in a biologically relevant context with a long term goal of building a functional map of human genome. Introduction of small interfering RNAs (siRNAs) into cells with transfection reagents results in efficient gene silencing. siRNA-based functional genomics is widely used in established cell lines in vitro, but its applicability to primary cells and in vivo target validatin has been limited because of lack of efficient and non-toxic delivery systems. In collaboration with RXi pharmaceuticals we have developed a novel class of RNAi compounds - self deliverable RNAs. These small, asymmetric, hydrophobically modified RNA compounds enter cells and tissues without requirement for delivery formulation and efficiently silence genes in vitro and in vivo, enabling functional genomics studies in primary cells, embryonic cells, tissues, ex vivo, and in vivo. The major technical hurdle, which impedes wide spread use of this platform by scientific community is the complex and costly process of compound identification, synthesis and validation. The focus of this fast-track proposal is optimization of compound discovery process (Phase I) and development of a panel of 200-1000 functionally validated sdRNAs (Phase II), Completion of this proposal will build a commercially available product platform that will revolutionize functional genomics studies in biologically relevant systems such as primary cells, stem cells, tissue and organ models and eventually in vivo.

描述（由申请人提供）：HGP的完成（人类基因组计划）已在生物医学研究中打开了“基因组时代”，并可以对各种基因元素进行功能基因组学分析。编码项目的长期目标是识别并在功能上表征所有基于序列的基因组元素。在编码中研究的功能元素包括转录序列，转录调节因子和RNA转录本身的调节剂。超过50％的转录基因序列的功能尚不清楚。它们中的大多数在专门的细胞中表达，因此只能在生物学相关的环境（即主细胞，体内）中研究其功能。在这一建议中，我们计划开发和商业化一个新的RNAi平台，该平台能够在生物学相关的环境中具有长期目标，即建立人类基因组的功能图。将小型干扰RNA（siRNA）引入具有转染试剂的细胞中，导致有效的基因沉默。基于siRNA的功能基因组学在体外已广泛用于已建立的细胞系中，但是由于缺乏有效且无毒的递送系统，其对原代细胞和体内靶标的验证蛋白的适用性受到限制。与RXI药品合作，我们开发了一种新型的RNAi化合物 - 自交付RNA。这些小的，不对称的，疏水的RNA化合物进入细胞和组织，无需递送配方，并有效地在体外和体内沉默基因，从而在原代细胞，胚胎细胞，组织，Ex Vivo和体内促进功能基因组学研究。科学界阻碍了该平台广泛使用该平台的主要技术障碍是复合识别，合成和验证的复杂且昂贵的过程。这项快速轨道提案的重点是优化复合发现过程（I阶段）和200-1000个经过功能验证的SDRNA（II阶段）的小组的开发，该提案的完成将建立一个市售的产品平台，该平台将彻底改变生物学相关的系统中的功能基因组学研究，例如主细胞，干细胞，组织，组织和器官和最终In Inivo。