Utilization of CELL PICKER and MCE-202 MultiNA in the eld of genome editing

Importance of the Topic

The rapid adoption of genome editing techniques such as CRISPR/Cas9 has transformed multiple areas of research and industry, including animal breeding, regenerative medicine, and molecular diagnostics. By enabling precise insertions, deletions, or replacements at defined genomic loci, these tools outperform earlier methods based on random recombination, offering higher efficiency and fidelity.

Objectives and Study Overview

This report illustrates how integration of automation and high-throughput electrophoresis can streamline the downstream cloning and validation steps following genome editing. Specifically, it evaluates Shimadzu’s CELL PICKER for colony separation and MCE-202 MultiNA for mutation confirmation in a workflow aimed at isolating and verifying edited cell clones.

Used Instrumentation

• CELL PICKER: Automated cell colony picking and transfer system with integrated microscope and image capture.
• MCE-202 MultiNA: Microchip electrophoresis platform for sizing and quantifying DNA/RNA fragments.

Methodology

The validated workflow comprises six key steps: (1) genome editing of target cells, (2A) cloning via colony separation or (2B) limiting dilution, (3) culturing cell clones, (4) mutation confirmation, (5) sequence verification, and (6) final clone isolation. CELL PICKER automates colony selection and transfer in step 2A, while MCE-202 MultiNA performs heteroduplex mobility assays in step 4 to detect small indels.

Main Results and Discussion

Integration of the CELL PICKER reduced per-colony handling time to approximately 2 minutes, cutting total labor by half compared to manual picking and eliminating user-to-user variability. Time-lapse imaging and immunostaining of picked induced pluripotent stem cell colonies confirmed viability and undifferentiated marker expression over six days.

Using MCE-202 MultiNA for heteroduplex mobility assays, PCR amplicons from edited genomic regions were heat-denatured and reannealed to form homoduplexes and heteroduplexes. Electrophoretic separation on microchips readily distinguished mutation-bearing samples by altered migration profiles, enabling rapid prescreening prior to sequencing.

Benefits and Practical Applications

• Enhanced throughput and consistency in colony isolation, reducing operator dependency and error.
• Objective, numeric detection of small indels without immediate sequencing, streamlining candidate selection.
• Broad applicability across gene-editing projects in research, biopharmaceutical development, and quality control laboratories.

Future Trends and Opportunities

Advances in automation and microfluidics are expected to further integrate genome editing workflows, enabling fully automated cell line development pipelines. Combining high-content imaging, robotics, and microchip electrophoresis could accelerate therapeutic cell-line generation and precision breeding programs. Future enhancements may include AI-driven colony selection and multiplexed mutation analysis on single chips.

Conclusion

By incorporating CELL PICKER and MCE-202 MultiNA, laboratories can significantly improve the efficiency and reliability of post-editing cell cloning and screening. This integrated approach reduces manual labor, enhances reproducibility, and provides rapid, cost-effective mutation confirmation, addressing critical bottlenecks in genome editing workflows.