I am a beginner of CRISPR pooled screen. I read the paper titled "Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening" (Julia Joung et al., 2017). In the paper, for a library size of 100,000 unique sgRNAs, we transduce 1.67 × 10^8 cells at an MOI of 0.3 at the start. After puromycin addition, I need...
I am currently working on a CRISPR screen using the Brunello Library, 2-vector system. The Cas9 portion contains Blasticidin selection and the sgRNA library has puromycin selection. After infection and antibiotic selection (with both Blast and Puro), we did several rounds of FACS. However, after sorting, we found that thos...
I'm going to use gecko library in order to infect primary macrophages and is the very first time I'm approaching this technique, so I have some questions. I'm following the nat prot paper by Julia Joung (2017) but I have some questions regarding the titration part. I'm using the 2 vectors system and I will infect macropha...
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