A lectin-based magnetic CRISPR screening protocol permits rapid discovery of genes regulating cell-surface glycosylation

Kim, J.; Kovacs, H.; Wisnovsky, S.

doi:10.1101/2025.09.15.676433

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bioRxiv · 10.1101/2025.09.15.676433

A lectin-based magnetic CRISPR screening protocol permits rapid discovery of genes regulating cell-surface glycosylation

Kim, J.·Kovacs, H.·Wisnovsky, S.

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Abstract

FACS-based CRISPR screening has emerged as a potent tool for dissecting the genetic networks that regulate cell-surface glycosylation. However, existing protocols are tedious and poorly suited to many cell models. We developed a lectin-based magnetic-activated cell sorting platform (Lec-MACS) that enables rapid identification of genes controlling expression of specific cell-surface glycans. Lec-MACS is faster and easier to perform than FACS-based screening while producing data of similar quality. We subsequently applied Lec-MACS to produce a genomic atlas of genes regulating breast cancer hypersialylation. This method will dramatically expand the scope and throughput of genetic screens targeted at cell-surface glycans.

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Kim, J., Kovacs, H., Wisnovsky, S.. 2025-09-16. A lectin-based magnetic CRISPR screening protocol permits rapid discovery of genes regulating cell-surface glycosylation. https://doi.org/10.1101/2025.09.15.676433

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