Convenient and Quick Isolation and Analysis of Genome-Edited Cells Using PERFLOW Sort and MultiNA

Introduction

With the growing popularity of genome editing techniques, a major concern for researchers is: how efficiently intended edits can be achieved on cells or organisms, and how the type of editing made can be identified by a high-throughput process. These processes of concern, however, are described in papers as if they are very simple procedures. For example, in a paper which describes established genome-edited cells and their characterization analysis, the establishment of a mutated cell line is often summarized in a sentence like the one indicated below (the description about the transfection amount and incubation time is omitted). "Genome-editing tools were delivered by the ** method, and single-cell cloning was performed to obtain ** and ** clones of heterozygously and homozygously mutated cells, respectively." Unless it is a technological treatise, the data presented for this process would be the genotyping results of the several cell lines which were used as target mutant cells in the final analysis. A laboratory leader who has read such a paper may without careful consideration instruct a researcher to create cells by inducing mutations into the ** gene in the same way as in this paper. However, the cell line and gene delivery method to use may not be the same, and even if they are the same, depending on the target gene, the targeting efficiency changes and there is no guarantee that the same mutant cell line can be established. Furthermore, the process of single-cell cloning or mutant cell screening involves complicated procedures which do not appear in papers. This Application Note introduces a pipeline for establishing genome-edited cells efficiently together with an actual example to help researchers resolve such working-level concerns.

February 5, 2019 GMT