PinPoint-FC System for Platform Cell Line Generation & Retargeting (includes PIN300A-1, FC200PA-1, PIN200A-1, PIN510A-1, & PIN600A-1)

Efficient Isogenic Cell Line Generation and Retargeting

Product Description

With the PinPoint Targeted Integration System, you can ensure that only a single copy of your transgene is inserted into the genome.

• Create isogenic cell lines
• Achieve high-efficiency integration with no insert size-limit
• Turn cell line construction into a high-throughput process

Making cell line generation and retargeting reliable, efficient, and amenable to high-throughput production, the PinPoint-FC System for Platform Cell Line Generation and Retargeting provides all the plasmids you need for implementing the PinPoint System using the PhiC31 Integrase to place the PinPoint attP site into the target genome.

Learn more about Targeted integration with the PinPoint System

• PinPoint-FC attP Placement Vector (# PIN300A-1)
• PhiC31 Integrase Expression Plasmid (# FC200PA-1)
• PinPoint Integrase Expression Plasmid (# PIN200A-1)
• CAG PinPoint Donor Vector (# PIN510A-1)
• CAG-GFP-T2A-Luc PinPoint Positive Control Donor Vector (# PIN600A-1)

Supporting Data

Insulated PinPoint Placement Vector colony count assay

Figure 1. Efficient placement of the PinPoint attP site using the PhiC31 System. Step 1 of the PinPoint Targeted Integration System—the PinPoint attP site was placed into HEK293 cells using the PinPoint-FC Placement Vector (Cat.# PIN300A-1) and the PhiC31 Integrase (Cat.# FC200PA-1). Positive cells were selected with G418 (neomycin resistance) for fourteen days and six separate lines were picked and expanded. After eleven days, the cells were fixed and stained with a solution of 50% methanol plus 1% methylene blue. The plates were washed twice with PBS and allowed to air dry. Only the plate of cells that were transfected with the PhiC31 Integrase Expression Plasmid showed a robust number of cells.

Puromycin-Resistant HEK293T Colonies after Targeting with PIN600A-1-SBI

Figure 2. Targeted integration with the PinPoint Integrase and a PinPoint Donor Plasmid. Step 2 of the PinPoint Targeted Integration System—the cell lines created in Figure 1 were transfected with a Positive Control PinPoint Gene Donor Vector expressing GFP (Cat.# PIN600A-1) for precise integration. Unlike the R4 integrase used in a similar system, the PinPoint Integrase does not recognize pseudo-sites and will only integrate at its placed recognition sequence. This feature results in the increased efficiency of correctly retargeted cell lines compared to the R4 integrase system. The entire therapeutic gene expression cassette is active and fully insulated with cHS4 DNA elements.