GSM8720911: single-cell multiome datasets (ATAC + Gene Expression), sgRNA enrichment, CRISPRi; Homo sapiens; RNA-Seq
Adherent cells were digested with 0.25% trypsin and collected in a 15mL tube containing serum-containing medium, followed by centrifugation and washing of the cell pellet with serum-free basal medium to obtain a single-cell suspension. The library construction protocol for single-cell multiome datasets (ATAC + Gene Expression)(scMulti) was performed following the manufacturer's instructions using the Chromium Single Cell Multiome ATAC + Gene Expression Reagent Kits. Nuclei are resuspended and transposed with a Transposase to fragment DNA in open chromatin regions and add adapter sequences. The transposed nuclei are then encapsulated into Gel Beads-in-Emulsion (GEMs) with barcoded primers for both ATAC and gene expression. After GEMs are broken, the products are purified and amplified via PCR to generate sufficient material for library construction. The final ATAC and gene expression libraries are constructed by adding sample indices, size selection, and adapter ligation. Sequencing is performed on an Illumina platform with paired-end reads, capturing both ATAC and gene expression data, including barcodes, UMIs, and sample indices. Finally, the enriched sgRNA library and fragmented cDNA were sequenced on an Illumina NovaSeq6000 PE150 platform, following the standard 10x libraries configuration.