10k A375 Cells Transduced with (1) Non-Target and (1) Target sgRNA, Single Indexed

10K A375 cells, a human cell line derived from malignant melanoma cells, were transduced with non-target and target sgRNA. A375 dCas9 Parental Cells were transfected separately with individual guides (either non-targeting or Rab1a sgRNA, with Capture Sequence 2 inserted into the hairpin structure of the sgRNA). Selected cells (cultured in a selection media) for each condition were individually frozen. Aliquots of Rab1a and non-target sgRNA containing cells are thawed, counted, and mixed 1:1. The same cells were used to generate Chromium Single Cell 3’ (Dual Index) data.

Libraries were prepared following the Chromium Single Cell 3' Reagent Kits v3.1 User Guide with Feature Barcode technology for CRISPR Screening (CG000205) and sequenced on Illumina NovaSeq 6000.

Single Cell 3’ CRISPR Screening v3.1 Single Index Library

• Sequencing Depth: 81,078 read pairs per cell

• Paired-end, single indexing Read 1: 28 cycles (16 bp barcode, 12 bp UMI) i7 index: 8 cycles (sample index) Read 2: 91 cycles (transcript)

• Analysis Parameters Used: Run with --expect-cells=10,000

Key metrics were:

• Cells detected: 11,503
• Median UMI counts per cell: 32,640
• Median genes per cell: 5,721

This dataset is licensed under the Creative Commons Attribution license.