Journal: bioRxiv

Article Title: CRISPR–Cas9 Editing of Myeloid Lectins Reveals Target-Dependent Transcriptional Remodeling in Tumor-Associated Macrophages

doi: 10.64898/2026.10.02.756241

Figure Lengend Snippet: Screening and selection of sgRNAs targeting candidate lectins in HeLa S3 cells. A. Schematic of the sgRNA screening workflow. HeLa S3 cells were nucleofected with pre-assembled Cas9–sgRNA RNPs targeting CLEC4E, CLEC6A, CD209, SELL, or SIGLEC9, with CD14 included as a technical reference control. Genomic editing was assessed three days after nucleofection by Sanger sequencing and ICE analysis to support selection of sgRNAs for subsequent evaluation in primary human monocytes. B. Schematic representation of the lectins genomic locus showing annotated RefSeq transcript isoforms, exon organization (blue), and the position of the targeting sgRNAs (orange). C. Predicted frameshift frequencies determined by ICE analysis following individual sgRNA delivery in HeLa S3 cells. Hatched bars indicate the sgRNA selected for each target for subsequent evaluation in primary human monocytes. Data represent a single technical screening experiment.

Article Snippet: BT474 and HeLa S3 cell lines were acquired from ATCC and cultured in RPMI 1640 or DMEM medium (Gibco), respectively, both supplemented with 10% (v/v) FBS (Gibco).

Techniques: Screening, Selection, Control, DNA Sequencing, Analysis