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

Significance of the Topic

Genome editing via CRISPR/Cas9 and related nucleases has revolutionized genetic modification by enabling targeted, efficient alterations in diverse organisms. Automating labor-intensive steps like colony picking and mutation confirmation addresses throughput and reproducibility challenges in research and industrial settings.

Objectives and Overview of the Study

This work demonstrates the integration of Shimadzu's CELL PICKER and MCE-202 MultiNA systems into a genome editing workflow to enhance the efficiency of isolating and verifying clones carrying desired edits. The focus is on automating colony separation and confirming mutations to accelerate downstream analysis.

Methodology and Instrumentation

The standard genome editing workflow comprises six stages: gene modification (via CRISPR/Cas9 or other nucleases), clone isolation by colony separation or limiting dilution, culture expansion, mutation confirmation, gene verification, and final clone recovery. CELL PICKER automates stage two (colony separation) by selecting and transferring individual colonies under microscope control. MCE-202 MultiNA implements heteroduplex mobility assays at stage four, using microchip electrophoresis to detect insertions or deletions by differences in DNA strand migration.

Instrumentation Used

• CELL PICKER: automated colony picking tool with pipetting unit, integrated microscope, and camera.
• MCE-202 MultiNA: fully automated DNA/RNA microchip electrophoresis system enabling high-throughput sizing and quantification.

Main Results and Discussion

In benchmarking, CELL PICKER reduced the total time for picking 48 colonies by over 50%, maintaining a consistent rate of under two minutes per colony compared to manual methods. Time-lapse imaging and immunostaining of picked iPS cell colonies confirmed normal proliferation and retention of pluripotency markers post-picking. The MCE-202 MultiNA heteroduplex mobility assay produced clear, reproducible patterns distinguishing wild-type, mutant, and heteroduplex DNA species, facilitating rapid pre-screening of edited clones before sequencing.

Benefits and Practical Applications of the Method

• Enhanced throughput and reproducibility in clone isolation.
• Reduced operator variability and skill dependency.
• Cost-effective mutation pre-screening to prioritize samples for sequencing.
• Wide applicability in academic research, biopharmaceutical development, quality control, and synthetic biology.

Future Trends and Potential Applications

Advances may include integration with machine learning–driven colony selection, coupling with high-content imaging for phenotypic screening, adaptation to single-cell editing platforms, and incorporation into automated end-to-end genome engineering pipelines for precision medicine and industrial biotechnology.

Conclusion

The combined use of CELL PICKER and MCE-202 MultiNA significantly streamlines the post-editing workflow by automating colony picking and delivering rapid, reliable mutation screening. This integrated solution enhances efficiency, consistency, and scalability in generating validated genome-edited cell lines.