We narrowed to 3 results for: cas9 expression plasmid

1. Isolating a Monoclonal Cell Population by Limiting Dilution

   Type

   Protocol

   ...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...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...sufficiently expanded, screen the lines for transgene expression and/or other phenotypes. For example, perform...

2. Virus Protocol - Generating Stable Cell Lines

   Type

   Protocol

   ...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...

3. Lentivirus Production

   Type

   Protocol

   ...DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your insert of interest... cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ... 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios...