...individual A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast 1 and then selected...Figure 2: Cas9 expression in monoclonal cell lines generated from A549 cells transduced with lentiCas9-Blast... not express Cas9 and are included as a control (WT). Vinculin is a loading control. 1 lentiCas9-Blast...could lead to reduced transgene expression over time, as the lower expressing clones take over the polyclonal...Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening...that are more likely to retain stable transgene expression. Other methods of generating a monoclonal cell...population be monitored regularly for transgene expression. This protocol has been optimized for adherent...

...individual A549 cells stably expressing Cas9. A549 cells were transduced with lentiCas9-Blast and then selected... the most desirable level of expression. Over time, transgene expression in a polyclonal population may...single cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described...Protocol to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector...protocol can be used to generate stable cell lines expressing a gene of interest from an integrated lentiviral...lentiviral vector. Unlike the short-term protein expression observed using transient transfection approaches... lentiviral vectors enables long-term protein expression studies. Moreover, repeating experiments in a...