human crispr brunello lentiviral pooled library  (Addgene inc)

Journal: bioRxiv

Article Title: High-throughput Genome Wide CRISPR Knock Out mechanical sort identifies genes driving metastatic cancer cell softening

doi: 10.64898/2026.02.12.705447

Figure Lengend Snippet: Cancer cells expressing Cas9 are transduced with a lentiviral library containing sgRNAs across the entire protein-coding genome at a low MOI to allow for single gene knockouts. Antibiotic selection ensures only cells with gene knockouts are included in mechanical screen. Next, the library of cells with single gene knockouts are processed with a microfluidic stiffness-based cell sorting device into 5 mechanical subsets. The inset shows a side and top view of the ridged region of the device where cells must deform underneath each diagonal ridge. Softer cells deform easily underneath the ridges and follow hydrodynamic streamlines to provide a slight negative deflection towards outlets 1 and 2. Stiffer cells resist deformation and are deflected along the diagonal ridge resulting in trajectories towards outlets 4 and 5. Genomic DNA is harvested from each mechanical subset, the sgRNA regions are amplified and sequenced, and the distribution of sgRNAs before and after the mechanical screen is used to determine the correlation of gene knockouts with mechanical subsets.

Article Snippet: The human CRISPR Brunello lentiviral pooled library was designed to optimize on-target activity and reduce off-target effects (Addgene 73178-LV) .

Techniques: Expressing, Transduction, Selection, FACS, Amplification

Journal: Cancer Science

Article Title: Development of a Synthetic Lethality‐Based Combination Therapy Using LIG1 and PARP Inhibitors for Prostate Cancer

doi: 10.1111/cas.70194

Figure Lengend Snippet: Genome‐wide CRISPR screening identifies synthetic lethal or resistant candidates with PARP inhibition in prostate cancer. (A) Genome‐wide CRISPR‐Cas9 knockout screen in DU145, 22Rv1, and LNCaP prostate cancer cell lines. Cells were transduced with a gRNA library targeting 18,010 genes and treated with olaparib or DMSO for 14 days before next‐generation sequencing analysis. (B) Dot plots showing negatively selected genes following olaparib treatment indicate potential synthetic lethal interactions with PARPIs. The names of the top 10 genes are shown. (C) Dot plots displaying positively selected genes after olaparib treatment that suggest potential resistance mechanisms to PARPIs. The names of the top 10 genes are shown.

Article Snippet: We used the Human Improved Genome‐wide Knockout CRISPR Library v1, containing 90,709 gRNAs targeting 18,010 genes, which was gifted by Kosuke Yusa (Addgene #67989) [ ].

Techniques: Genome Wide, CRISPR, Inhibition, Knock-Out, Transduction, Next-Generation Sequencing

Journal: medRxiv

Article Title: Modelling Acral Melanoma in Admixed Brazilians Uncovers Genomic Drivers and Targetable Pathways

doi: 10.1101/2025.08.08.25332963

Figure Lengend Snippet: A) Overview of the genome-wide CRISPR knockout screens performed in AM-PDX-derived cell lines. Scheme was produced using Biorender.com. B) Waterfall plot illustrating genes identified as common dependencies in AM007 and AM016 through BAGEL analysis, ranked by their scaled Log Fold Change (sLFC). Selected genes are shown. Dotted lines indicate a significant threshold (0.05 < sLFC < −0.05). C) Comparison of genes identified as essential in acral melanoma (AM, n=5 cell lines) and other tumour types (pan-cancer, n=1010 cell lines) using DepMap datasets (p < 0.05, Mann-Whitney U test). D) Venn diagram showing the overlap of specific dependencies identified in acral, cutaneous, and uveal melanoma, as described in panels C and D. E) Comparison between genes identified as essential in AM (n=5 cell lines) with genes in cutaneous (CM, n=56 cell lines) and uveal melanoma (UM, n=10 cell lines) using DepMap datasets (p < 0.05, Mann-Whitney U test). For panels C and D, only genes identified as more essential in AM are shown. Box plot features: centre line = median; box limits = upper and lower quartiles; whiskers = 1.5x interquartile range; points = outliers.

Article Snippet: The Human Improved Genome-wide Knockout CRISPR Library v1.1, a gift from Kosuke Yusa (Addgene, 67989), was packaged into lentivirus in HEK293T cells, and BFP expression was used for titration.

Techniques: Genome Wide, CRISPR, Knock-Out, Derivative Assay, Produced, Comparison, MANN-WHITNEY