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.
This project will develop will develop techniques for rapid screening of gene edited cells containing the correction insertion. These techniques will greatly facilitate biological and biomedical research in a wide range of fields by making gene editing of mammalian cell lines more efficient and accessible.
