...expression vector Cas enzyme promoter can be constitutive (CMV, EF1alpha, CBh) or inducible (Tet-ON); U6 ...efficiency Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector or separate...components to make lentiviral particles (for details about lentivirus, see our Lentivirus Guide ) Stable,...lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or...separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify and enrich...CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles (for details...oligos and clone these oligos into your desired vector. In many cases, targeting oligos are synthesized...

...pooled format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which...CRISPR libraries What Are Pooled Lentiviral CRISPR Libraries? Pooled lentiviral CRISPR libraries are a heterogeneous...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual...Cas9-based genome engineering from a single lentiviral vector. Nucleic Acids Research , 42 (19), e147. PMID...8E). In a 2-vector system, you’ll need to either co-infect with a Cas9-producing lentivirus or use a Cas9...positively-charged grooves on Cas9. gRNA binding induces a conformational change in Cas9, shifting it into...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...

...Commonly used cloning vectors include Gateway entry vectors and TOPO cloning vectors. If you are looking...combination of a DNA fragment with a plasmid or vector DNA backbone generates a recombinant DNA molecule...specifically engineered plasmids, commonly referred to as vectors, which have become possibly the most ubiquitous...facilitate the cloning of DNA fragments. Cloning vectors tend to be very simple, often containing only a...expression (for the purposes of gene study). Expression vectors must contain a promoter sequence, a transcription...the transcription process to stop. An expression vector can also include an enhancer sequence which increases... amount of protein or RNA produced. Expression vectors can drive expression in various cell types (mammalian...