Rapid screening of gene-edited cells

项目摘要 最近，基因编辑技术的突破使生物和生物医学的许多领域发生了革命性的变化 研究，支持包括大规模标记内源基因或编辑非编码在内的应用 基因组元素。然而，对于那些包含正确编辑的编辑细胞系的筛选，主要有 依赖于经典的克隆选择方法，这种方法速度慢，资源密集。在此，我们建议 开发活细胞内的酶扩增技术以报告1)低丰度的表达 蛋白质和2)基因组中特定DNA序列的单一拷贝。这种技术将使快速隔离成为可能 通过简单的基于荧光的细胞分选来编辑细胞，从而大大提高了效率和可及性 关于细胞系的基因编辑。

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.