genome wide crispr sgrna library  (Addgene inc)

Journal: bioRxiv

Article Title: Antibody-Dependent Heterotypic Syncytia Drive COVID-19 Inflammation and Disease Progression

doi: 10.64898/2026.02.11.705426

Figure Lengend Snippet: A , Schematic of the CRISPR-Cas9 screen. B , Volcano plot of genes from the screen, ranked by fold change and adjusted P value. C , Representative flow cytometry dot plots of GW01-induced syncytia between A549-spike and FCER1G -KO (FcRγ-KO) THP-1 clones versus non-targeting control (NT), and quantification of syncytia(right inset)(mean ± SEM, n=3, * P <0.05, **** P <0.0001). D , Representative plots A549-spike fusion with ADAM10-KO THP-1 clones versus control and quantification of syncytia. E , Rescue of FcRγ: representative plots for FcRγ-KO-LVX (empty lentiviral vector), FcRγ-KO-FcRγ (FcRγ cDNA), and NT-LVX; with quantification of syncythia. F , Rescue of ADAM10: representative plots for ADAM10-KO-LVX (empty vector), ADAM10-KO-ADAM10 (ADAM10 cDNA), and NT-LVX, along with quantification of syncytia. G , Effect of CD64 or CD32 blocking antibodies on GW01-dependent syncytium formation measured by NanoLuc assay. H , Effect of the ADAM10 inhibitor GI254023X on GW01-dependent syncytium formation. I , Representative fluorescence images of syncytia (yellow merge) between THP-1-mCherry (red) and A549-spike (green; CFSE) at 3 hour post co-culture, with or without GI254023X. Scale bar, 100 μm. Data are presented as mean ± SEM, n=3, * P <0.05, *** P <0.001, **** P <0.0001 by one-way ANOVA.

Article Snippet: The genome-wide CRISPR sgRNA library (Addgene, 101926-101934) was packaged into lentiviruses.

Techniques: CRISPR, Flow Cytometry, Clone Assay, Control, Plasmid Preparation, Blocking Assay, Fluorescence, Co-Culture Assay

Journal: Cell Reports Medicine

Article Title: CDK4/6 inhibition overcomes venetoclax resistance mechanisms with enhanced combination activity in acute myeloid leukemia

doi: 10.1016/j.xcrm.2025.102526

Figure Lengend Snippet: Combining ven+palbo mitigates single-agent resistance due to clinically observed mutations (A) Enrichment of individual sgRNAs for RB1, BAX, and IKZF1 shown as fold change over DMSO control following a 21-day exposure to palbo, ven, or ven+palbo in OCI-AML2 Cas9 C6 cells. (B) Immunoblot showing efficiency of knockdown of RB1, BAX, and IKZF1 proteins in OCI-AML2 cell lines. A cell line expressing an NT sgRNA was used to generate a control cell line. Vinculin was used as a protein loading control. Par, parental; NT, non-targeting. (C–F) Dose-response curves for OCI-AML2 NT and KO cell lines evaluated for drug sensitivity to palbo, ven, or the combination. Data points denote the mean normalized cell viability ± SD for 3 replicates. (G) IC 50 values derived from dose-response curves of OCI-AML2 cell line drug sensitivity assays shown in (C)–(F). Data represent the mean IC 50 ± SD for 3 replicates (∗ p ≤ 0.05 and ∗∗ p ≤ 0.01 by Student’s t test). (H–J) Outgrowth of OCI-AML2 Non-targeting (H), OCI-AML2 Bax KO (I), and OCI-AML3 cell lines (J) treated with palbo, aza, and ven single agents, duplicate combinations and the triplet. Total viable cells over a 14-day drug treatment are shown. Data points denote the mean total number of viable cells ± SD for 3 replicates. One-way ANOVA with Tukey’s post-test for multiple comparisons was used for day 7 and day 14 time points as indicated. (∗ p ≤ 0.05, ∗∗ p ≤ 0.01, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001) (K) Immunoblot of apoptotic proteins in OCI-AML2 cells, drug treated for 14 days.

Article Snippet: Clonal OCI-AML2 Cas9 C6 cells were used for genome wide knockout with sgRNA library (Addgene #67989) as described above.

Techniques: Control, Western Blot, Knockdown, Expressing, Derivative Assay

Journal: Cancer Science

Article Title: In‐Tumor CRISPR ‐Cas9 Knockout Screening and Novel Therapy Development for Malignant Transformation of Ovarian Teratoma

doi: 10.1111/cas.70315

Figure Lengend Snippet: Integrated CRISPR‐Cas9 screening identifies key genes for tumor growth and cisplatin resistance in MTMCT. (A) Schematic overview of in vivo CRISPR‐Cas9 knockout screening. NOSCC1 cells transduced with the sgRNA library were divided into two groups—in vitro‐cultured reference cells and in vivo mouse tumors. (B) Scatter plot of negatively selected sgRNAs from in vivo tumors, showing fold‐change versus baseline gene expression (TPM) in NOSCC1. Red dots highlight the genes with TPM > 100. (C) Scatter plot of negatively selected sgRNAs from CDDP‐treated cells, showing fold‐change versus baseline gene expression (TPM) in NOSCC1. Red dots highlight the genes with TPM > 100. (D) Workflow for candidate gene prioritization and integration of transcriptomic and spatial transcriptomic data.

Article Snippet: Genome editing was performed using the Guide‐it CRISPR Genome‐Wide sgRNA Library System (Takara Bio, Japan), following the manufacturer's protocol.

Techniques: CRISPR, In Vivo, Knock-Out, Transduction, In Vitro, Cell Culture, Gene Expression