CRISPR/CAS DROPOUT SCREENING PLATFORM TO REVEAL GENETIC VULNERABILITIES ASSOCIATED WITH TAU AGGREGATIONCas-protein-ready tau bio sensor cells, CRISPR/Cas synergistic activation mediator (SAM)-ready tau biosensor cells, and methods of making and using such cells to screen for genetic vulnerability ...
To test the generalized suitability of T.gonfio-mediated epigenetic silencing, we benchmarked T.gonfio against a refined pooled dual-sgRNA CRISPRoff library, which prioritizes guides according to a more expansive set of screening data58,60. The TagBFP-expressing qgRNA T.gonfio (pool of 22,...
1c). We investigated if low amino acid conditions altered transporter gene expression; low-Arg and low-Leu screening conditions increased expression of most genes in our SLC7 transporter panel, but not all of them, consistent with results for HEK293T cells in single amino acid dropout media35...
Here we performed a genome-wide CRISPR-Cas9 loss-of-function screen in MCF-7 ER+ BC cells, executed under pharmacological pressure with SI-12, to identify targets whose inhibition will enhance SI-12 anti-tumor activity. Identification of genes whose “dropout” associates with increased sensitivity...
Prior to the screening, Cas9/enCas12a-expressing cell lines were selected with blasticidin then transduced with the CRISPR library viruses in three biological replicates to achieve a representation of 750 cells per sgRNA and at low multiplicity of infection (MOI; 0.3–0.5). Cell lines were ...
Pooled CRISPR screening data in cancer cell lines is limited to Log2(fold changes) for PMAIP1 sgRNAs and is available upon request. RNA-seq data processing and pooled CRISPR NGS were conducted using open source software as indicated in the key resources table. Acknowledgments We would like ...
These observations argue for the devel- opment of methods such as CRISPRcleanR, which are in- dependent of CN values for the analysis of CRISPR-KO screening data, and indicate that biased responses are not solely due to the amount of DNA damage and may also be caused by additional factors...
they require contrasting one subset of the data with another, but they can yield spurious results if the “test” and “control” subsets are not adequately matched in other respects. Thus, rigorous testing frameworks are needed that efficiently make use of the data and control for such biases...
Next, we sought to understand how gRNA position within the CDS of the target gene impacts gRNA activity. Given that most gRNAs in Project Achilles were located in the first 50% of the target CDS by design, we obtained a different screening dataset; we downloaded data from a genome-wide fit...
Source data Supplementary Figure 5 A pilot screen to identify optimal conditions for UMI-based CRISPR screen analysis. Setup of screen; Upon editing, various clonal outgrowth regiments, followed by clonal expansion and dropout screening, were run in parallel. Cas9 expression was induced by Dox, ...