Semi-automated optimized method to isolate CRISPR/Cas9 edited human pluripotent stem cell clones.

ABSTRACT:

Background

CRISPR/Cas9 editing systems are currently used to generate mutations in a particular gene to mimic a genetic disorder in vitro. Such "disease in a dish" models based on human pluripotent stem cells (hPSCs) offer the opportunity to have access to virtually all cell types of the human body. However, the generation of mutated hPSCs remains fastidious. Current CRISPR/Cas9 editing approaches lead to a mixed cell population containing simultaneously non-edited and a variety of edited cells. These edited hPSCs need therefore to be isolated through manual dilution cloning, which is time-consuming, labor intensive and tedious.

Methods

Following CRISPR/Cas9 edition, we obtained a mixed cell population with various edited cells. We then used a semi-automated robotic platform

SUBMITTER: Frank E 

PROVIDER: S-EPMC10142500 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature