...Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast 1 and then selected with...Figure 2: Cas9 expression in monoclonal cell lines generated from A549 cells transduced with lentiCas9-Blast... express Cas9 and are included as a control (WT). Vinculin is a loading control. 1 lentiCas9-Blast was...additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts... pool of stable cells. Transducing cells with lentivirus results in a heterogenous polyclonal population...protocol for generating stable cell lines with lentivirus . Day 0: (optional) Seed cells for the generation...

...A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected with...single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described... Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature...cells that have taken up the lentiviral transgene. Note that not all lentiviral vectors deliver antibiotic...Thaw the lentiviral aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM.... This is because lentiviral integration is random. Given the MOI of the lentivirus used, cells may also...an integrated lentiviral vector. Protocols...