CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) is the advanced genomic gene editing technology. A target’s sequence specific guide RNA molecule (gRNA) directs a cas endonuclease to the genomic DNA target sequence. Then, the Cas enzyme creates a double-strand break at the target sequence that can be repaired by either Non-Homologous End-Joining (NHEJ), which can result in insertion or deletions (InDels), or correction / Homology Directed Repair (HDR). InDels can disrupt expression of the target gene while repair by HDR, which requires the presence of a repair template, allows modification of the gene.
CAS9 Expression Cell Line as CRISPR Ready:
Cas9 is the most frequently used cas endonuclease so far. GenTarget’s CRISPR ready cell lines are transformed from lentivirus transduction, expressing a standalone cas9 enzyme. Each cell line expresses the nuclear penetrating humanized wild-type Cas9 endonuclease (Streptococcus pyogenes).
The Cas9 enzyme is driven by our engineered super strong CMV promoter (suCMV). Each cell line also contains an antibiotic selection marker or an antibiotic-fluorescent fusion dual marker (GFP-puromycin or RFP-Puromycin) under the RSV promoter. The following expression cassette was integrated into each cell line’s genome.
Why use CRISPR Ready Cell Line:
The CRISPR ready (Cas9 expression) stable cell lines make the genomic editing easier than ever. You simply deliver the target specific gRNA into the CRISPR ready cell line and select the knock-out or knock-in cells for your desired target. No need to do cas9 containing CRISPR cloning, and no worry about the hard-to-delivered Cas9 constructs. You only need to generate the target-gRNA constructs which is much easier in cloning, and much smaller in size for delivery.
GenTarget also provide services to generate your target specific, ready-to-use gRNA lentivirus. Please Contact us if you need the CRISPR gRNA lentivirus services. Click Product Manual for product details.