Target site placement with the PinPoint-FC system

Platform cell lines containing the PinPoint attP site can be made through two different approaches. The PinPoint-FC system utilizes the phiC31 integrase (Cat# FC200A-1) along with the PinPoint-FC attP Placement Vector (Cat# PIN300A-1). This will integrate the cassette at pseudo attP sites within the targeted genome with typically single copy insertions.

Target site placement with the PinPoint-HR system

Alternatively, the PinPoint-HR system uses a site-directed placement strategy. Customized placement can be achieved using the PIN400A-1 HR vector. First, clone the homology arms for the desired integration site into the 5' and 3' MCS features. Then use this vector in combination with matched TALENs or Cas9 nuclease vectors. For more on assembling TALENs, click here. To learn more about building Cas9 vectors, click here. SBI has created a safe-harbor PinPoint placement vector (Cat# PIN410A-1) to insert the attP site at the human AAVS1 genomic locus for routine integration of gene expression cassettes at this site.