Automated Cloning for CRISPR Workflows Using the CellRaft AIR System

Gene editing workflows require the transformation of a large number of cells, followed by the isolation of individual cells from the larger population to establish clonal colonies. Given the large number of gene edits required for contemporary research and the labor-intensive components of the workflow, there is an unmet need to automate post-transformation cloning.

In this RaftNote we present data and protocols describing the CellRaft AIR™ System as a fully automated cloning platform. Using an imaging-based sorting modality, single cells can be monitored for transformation-positive phenotypes as well as their retention of clonality as colonies begin to grow. Using the protocols described here, we also provide evidence of improved cell viability with relatively robust HEK293 cells showing a nearly 3-fold improvement in viability.

Other laboratories have shown even more dramatic improvements in viability for more sensitive cells such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs). Finally, the system generates clonal colonies in 4-5 days, a significant improvement over the months required in more traditional workflows.