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Analyzing genomic elements in live animals by CRISPR imaging

通过 CRISPR 成像分析活体动物的基因组元件

基本信息

批准号：
8934113
负责人：
Bo Huang
金额：
$ 39.63万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-25 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8934113
关键词：
Adverse effects Age Aging Aneuploidy Animal Model Animals Base Pairing Benchmarking Binding Biological Models Biology Caenorhabditis elegans Cardiac Myocytes Cell Line Cells Chromosome Arm Chromosome Segregation Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complex DNA Sequence Detection Development Disease model Drug resistance Elements Event Fluorescent in Situ Hybridization Gene Dosage Gene Duplication Generic Drugs Genes Genetic Genome Genome engineering Genomic Instability Genomics Germ Lines Guide RNA HIV Infections Health Hepatocyte Heterogeneity Image Imaging Techniques Imaging technology Immune system Indium Individual Label Length Life Location Longevity Malignant Neoplasms Mammalian Cell Measurement Measures Megakaryocytes Methods Microfluidics Microscopy Mitosis Modeling Monitor Names Nematoda Optics Organism Play Polyploidy Process Proteins Recruitment Activity Repetitive Sequence Research Research Project Grants Retroviridae Sensitivity and Specificity Signal Transduction Sister Chromatid Stem cells Structure System Tandem Repeat Sequences Techniques Technology Telomerase Telomere Shortening Testing Time Work base cohesion design genetic element human disease imaging system impression improved insertion/deletion mutation movie nuclease programs single cell analysis single cell sequencing success targeted sequencing telomere tool

项目摘要

DESCRIPTION: Distinct from the common impression, the genomes of individual cells in our body are not exactly homogenous or static. Heterogeneity and dynamic changes occur in many cases, such as aging, cancer and HIV infection. Detecting these single cell events in a live animal remains a major challenge. In the proposed project, we plan to validate and further develop our recently invented CRISPR imaging technique in a multicellular organism. This technique utilizes the CRISPR/Cas9 system to fluorescently label specific, endogenous genomic loci for microscopy detection. Here, we will use the nematode C. elegans as the model organism to validate and benchmark CRISPR imaging as a technology to measure the size of repetitive genomic elements such as telomeres as well as the copy number of genes. We will also improve the sensitivity of CRISPR imaging for the detection of non-repetitive genomic elements. Finally, we will apply CRISPR imaging to the study of telomere length change at the single cell level during C. elegans development and aging.

描述：与普遍印象不同，我们体内各个细胞的基因组并不完全同质或静态。许多情况下都会发生异质性和动态变化，例如衰老、癌症和艾滋病毒感染。在活体动物中检测这些单细胞事件仍然是一个重大挑战。在拟议的项目中，我们计划在多细胞生物体中验证并进一步开发我们最近发明的 CRISPR 成像技术。该技术利用 CRISPR/Cas9 系统荧光标记特定的内源基因组位点以进行显微镜检测。在这里，我们将使用线虫秀丽隐杆线虫作为模式生物来验证和基准 CRISPR 成像技术，以测量端粒等重复基因组元件的大小以及基因的拷贝数。我们还将提高 CRISPR 成像检测非重复基因组元件的灵敏度。最后，我们将应用CRISPR成像来研究线虫发育和衰老过程中单细胞水平的端粒长度变化。