Analyzing genomic elements in live animals by CRISPR imaging

 描述：与通常的印象不同，我们身体中单个细胞的基因组并不完全是同源的或静态的。许多情况下都会发生异质性和动态变化，如老龄化、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和进一步开发我们最近发明的CRISPR成像技术。这项技术利用CRISPR/Cas9系统来荧光标记特定的内源基因组位点，用于显微镜检测。在这里，我们将使用线虫作为模式生物来验证和基准CRISPR成像作为一种测量重复基因组元件(如端粒)的大小以及基因拷贝数的技术。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成像技术在单细胞水平上研究线虫发育和衰老过程中端粒长度的变化。

 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.