CRISPR/Cas9 editing enables precise modifications in mesenchymal stem cells (MSCs), but isolating monoclonal clones remains a bottleneck. In this study, the F.SIGHT™ enabled gentle, fluorescence-guided single-cell isolation of MSCs edited to knock out RANKL, even when expression levels were low. With over 93% single-cell isolation efficiency, the system generated 182 viable clones – ultimately yielding two functionally validated RANKL-knockout lines capable of osteogenic differentiation.
These findings highlight the F.SIGHT as a high-throughput, gentle, and precise platform for generating CRISPR-edited monoclonal stem cell lines.
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F.SIGHT isolates CRISPR-edited MSCs with low fluorescent signals
Single-cell cloning achieved >93% efficiency, generating hundreds of viable clones
Functionally validated RANKL-knockout MSC clones were produced for regenerative applications