Rapid screening of gene-edited cells

项目摘要 最近，基因编辑技术的突破彻底改变了生物和生物医学的许多领域。 研究，使应用程序，包括大规模标记内源基因或编辑非编码 基因组元素然而，筛选包含正确编辑的细胞系， 依赖于经典的克隆选择方法，这是缓慢和资源密集型的。在此，我们建议 开发活细胞内的酶促扩增技术，以报告1）低丰度的 蛋白质和2）基因组中特定DNA序列的单拷贝。这项技术将使快速分离 通过简单的基于荧光的细胞分选， of gene基因editing编辑for cell细胞lines线.

Project Summary Recently breakthroughs in gene editing technology have revolutionized many fields of biological and biomedical research, enabling applications including large-scale tagging of endogenous genes or editing of non-coding genomic elements. Screening of edited cell lines for those containing the correct edits, however, have mostly relied on the classical clonal selection method, which is slow and resource-intensive. Here, we propose to develop an enzymatic amplification technique inside living cells to report 1) the expression of a low abundance protein and 2) a single copy of specific DNA sequence in the genome. This technique will enable rapid isolation of edited cells by simple fluorescence-based cell sorting, thus greatly enhancing the efficiency and accessibility of gene editing for cell lines.