... in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene (e.g. GFP...lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable, tunable expression ...transfer vector and used to generate AAV particles (for details, see our AAV Guide ) ∼4.5 kb packaging limit...Components of System Application Mammalian expression vector Cas enzyme promoter can be constitutive (CMV, EF1alpha...lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or... marker to identify and enrich positive cells Packaging and envelope plasmids provide the necessary components...oligos and clone these oligos into your desired vector. In many cases, targeting oligos are synthesized...

...format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included...need to generate lentivirus containing the entire library (Figure 8D). This lentivirus will then be used...8E). In a 2-vector system, you’ll need to either co-infect with a Cas9-producing lentivirus or use a Cas9...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR specificity...function. Its size allows it to be efficiently packaged into adeno-associated viruses (AAVs) . Other natural...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual...