lentiviral tkov3 sgrna library  (Addgene inc)

Journal: Nature cancer

Article Title: P2RY2-AKT activation is a therapeutically actionable consequence of XPO1 inhibition in acute myeloid leukemia

doi: 10.1038/s43018-022-00394-x

Figure Lengend Snippet: a) FACS-based CRISPR/Cas9 screening strategy to identify genetic modifiers of AKT phosphorylation in selinexor-treated AML cells. sgRNA library transduced OCI-AML2 cells were treated with selinexor for 48 hours, fixed/permeabilized and stained with phosphorylated AKT T308 primary antibody followed by Alexa Flour 488 conjugated secondary antibody. Stained cells were then sorted according to phosphorylated AKT T308 expression into high-expressing cells (top sort) and low-expressing cells (bottom sort). Genomic DNA was extracted and sgRNA barcodes were amplified and indexed prior to deep sequencing. The FACS screen gene score (FSGS) enumerates genes whose sgRNA representatives were enriched in the top or bottom sorted populations.

Article Snippet: . sgRNA library amplification: The Toronto Knockout CRISPR Library – Version 3 (TKOv3) was obtained from Addgene (Pooled Libraries #90294, #125517) and amplified according to provided published protocol.

Techniques: CRISPR, Phospho-proteomics, Staining, Expressing, Amplification, Sequencing

Journal: Nature cancer

Article Title: P2RY2-AKT activation is a therapeutically actionable consequence of XPO1 inhibition in acute myeloid leukemia

doi: 10.1038/s43018-022-00394-x

Figure Lengend Snippet: a) Scatterplot depicting gating strategy to isolate live sgRNA library transduced OCI-AML2 cells based on forward-scatter (FSC) and side-scatter (SSC).

Article Snippet: . sgRNA library amplification: The Toronto Knockout CRISPR Library – Version 3 (TKOv3) was obtained from Addgene (Pooled Libraries #90294, #125517) and amplified according to provided published protocol.

Techniques: CRISPR