human brunello kinome pooled library  (Synbio Technologies LLC)

Journal: bioRxiv

Article Title: Loss of JAK1 Function Causes G2/M Cell Cycle Defects Vulnerable to Kif18a Inhibition

doi: 10.1101/2025.02.19.638911

Figure Lengend Snippet: ( A ) Schematic of radiation CRISPR-Cas9 screening methodology. Cal27 and Detroit562 HSNCC cell lines were engineered to stably express Cas9, transduced with a kinome-wide gRNA library, and irradiated with 2 Gy for four consecutive treatments. Two weeks after treatment, gRNAs were isolated, PCR amplified, and sent for deep sequencing in parallel with unirradiated controls. Simplified screening results are shown for ( B ) Cal27 and ( C ) Detroit562 cell lines. Hits with an FDR ≤ 0.01 were considered significant. ( D ) Summary of pathways identified by the CRISPR screens including ( i) DNA repair, ( ii ) NFKB signaling, and ( iii ) JAK signaling.

Article Snippet: The Human Kinome CRISPR Knockout Library (Brunello) was purchased from Addgene (# 1000000082) and was generated as previously described .

Techniques: CRISPR, Stable Transfection, Transduction, Irradiation, Isolation, Amplification, Sequencing

Journal: Cancer Research Communications

Article Title: A Kinome-Wide Synthetic Lethal CRISPR/Cas9 Screen Reveals That mTOR Inhibition Prevents Adaptive Resistance to CDK4/CDK6 Blockade in HNSCC

doi: 10.1158/2767-9764.CRC-24-0247

Figure Lengend Snippet: CRISPR screening identified the cell-cycle pathway as the synthetic lethal pathway for mTORis in HNSCC. A, Scheme for CRISPR screening. Cal27-Cas9 cells were infected with the human kinome CRIPSR-knockout library, targeting 763 genes consisting of four sgRNAs for each gene, and subjected to vehicle treatment or INK128 treatment. At population doubling 20, genomic DNA was extracted from cells, and PCR and NGS were performed. B, Volcano plot of all dropout sgRNA hits. C, Extraction sequence of sensitizing sgRNA hits. The sensitizing hits were extracted with P value < 0.01 and analyzed with KEGG pathway analysis. D, Selected 109 hits plot. The hits included genes related to PI3K/mTOR and cell-cycle pathways. E, KEGG pathway analysis for sensitizing hits. KEGG pathway analysis was applied for significant 109 hits. The top 15 pathways are represented, and the color intensity of the bar represents the combined score. F, CCND1 mRNA expression and OS in TCGA-HNSCC patients. Patients with high ( z -score > 0.5, n = 124) and low ( Z -score < 0.5, n = 73) expression of each mRNA were compared using the log-rank test. G, CCND1 RPPA expression and OS in TCGA-HNSCC patients. Patients with high ( z -score > 0.5, n = 120) and low ( z -score < 0.5, n = 120) expression of each mRNA were compared using the log-rank test. (Created with BioRender.com .)

Article Snippet: Next, Cal27-Cas9 cells were infected with a human kinome CRISPR pooled library (Brunello, RRID: Addgene_75312) at representation of 650 and a multiplicity of infection of 0.3.

Techniques: CRISPR, Infection, Knock-Out, Extraction, Sequencing, Expressing

Journal: Cancer Research Communications

Article Title: A Kinome-Wide Synthetic Lethal CRISPR/Cas9 Screen Reveals That mTOR Inhibition Prevents Adaptive Resistance to CDK4/CDK6 Blockade in HNSCC

doi: 10.1158/2767-9764.crc-24-0247

Figure Lengend Snippet: FIGURE 1 CRISPR screening identified the cell-cycle pathway as the synthetic lethal pathway for mTORis in HNSCC. A, Scheme for CRISPR screening. Cal27-Cas9 cells were infected with the human kinome CRIPSR-knockout library, targeting 763 genes consisting of four sgRNAs for each gene, and subjected

Article Snippet: The human kinome CRISPR pooled library (Brunello) was a gift from John Doench and David Root (RRID: Addgene_75312).

Techniques: CRISPR, Infection, Knock-Out

Journal: Cell Reports Medicine

Article Title: Targeting of vulnerabilities of drug-tolerant persisters identified through functional genetics delays tumor relapse

doi: 10.1016/j.xcrm.2024.101471

Figure Lengend Snippet:

Article Snippet: Human Brunello kinome pooled library, guides 1–4 in lentiGuide-Puro backbone , Addgene , Cat# 75312.

Techniques: Recombinant, Membrane, cDNA Synthesis, Amplification, Mutagenesis, shRNA, Sequencing, Software