权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

High-precision pooled screening for quantitative molecular phenotypes

定量分子表型的高精度联合筛选

基本信息

批准号：
10462811
负责人：
NICHOLAS T INGOLIA
金额：
$ 30.66万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10462811
关键词：
Address Affect Bar Codes Biological Biological Process CRISPR screen CRISPR/Cas technology Cell physiology Cells Chromosome Mapping Complex DNA Data Dependence Engineering Event Gene Expression Gene Silencing Genes Genetic Genetic Epistasis Genetic Screening Genetic Transcription Genome Growth Guide RNA Human Individual Label Lead Learning Libraries Link Mammalian Cell Measurement Measures Molecular Monitor Mutagenesis Mutation Nucleotides Organism Outcome Pathway interactions Pattern Phenotype Population Population Sizes Process Proteins RNA Reagent Regulation Reporter Reporting Resolution Resources Saccharomycetales Signal Pathway Signal Transduction Sorting - Cell Movement Source Specific qualifier value System Techniques Transgenic Organisms Translating Translations cell growth computerized tools deep sequencing experimental study gene function genetic profiling genome-wide insight interest molecular phenotype novel precision genetics programs promoter response screening tool transcription factor

项目摘要

ABSTRACT The RNA-guided DNA recognition of CRISPR/Cas9 now enables comprehensive genetic screening in mammalian cells and other previously intractable systems. CRISPR/Cas9 provides highly programmable genetic perturbation through DNA cleavage, transcriptional inhibition, or targeted mutagenesis. Measuring the phenotypes associated with these genetic perturbations remains a challenge, and this often represents the greatest barrier to mapping the genetic dependencies of important biological processes. We propose to develop a general approach that links molecular phenotypes with the targeting guide RNAs that induce those effects in a large, pooled cell population. We label distinct guide RNAs with unique nucleotide barcodes that are expressed in an RNA reporter, linking the RNA abundance of the barcode to intracellular processes of interest. Our system can directly monitor transcriptional, post-transcriptional, and post-translational responses, allowing us to couple it with a wide array of intracellular signals. These expression measurements are well suited for epistasis analysis, which can identify genetic pathways and uncover novel gene functions through correlated patterns of genetic interaction from quantitative phenotypic proﬁles. We will make available our validated reagents for implementing this screening approach in budding yeast and in mammalian cells, providing a broadly useful resource for high-precision genetic proﬁling.

摘要