ABBIE (A Binding Based Integrase Enzyme)
The ABBIE system delivers consistent high integration efficiency across gene targets and cell types with minimization of off-target effects.
High Performance GLP/GMP Kits Coming Soon!
ABBIE System
The ABBIE system combines the ease-of-use site specificity of Cas9 protein with retroviral integrase driven integration of donor DNA sequences into genomic DNA.
This system provides guide RNA targeted integration of a desired DNA sequence (e.g. an expressable construct or gene-of-interest) without double stranded cutting of genomic DNA that would activate the cell’s DNA repair machinery. The system also avoids the use of viral vectors.
Cell Engineering
From using T-cells to fight cancer to using microbes to produce biofuels, cellular engineering is a growing technology enabling cutting edge breakthroughs across
science. Our world class team is developing proprietary cell lines leveraging the ABBIE technology. Please inquire regarding custom cell engineering projects.
Advantages of the ABBIE system
- Capable of integrating large DNA sequences (over 7 kb)into genomic DNA.
- Flexible system to alternate donor DNA sequence and targeting guide RNA.
- Provides faster time to a functional read out.
- No double cutting of genomic DNA.
Procedural Outline for ABBIE kit
Kit components- ABBIE recombinant protein; donor DNA vector with multiple cloning site; pre- tested control guide RNA; optimized buffers for pre-initiation complex formation and electroporation.
Steps;
1. Design and add donor polynucleotide sequences into provided donor construct.
2. Design guide RNA for targeting.
3. Combine ABBIE protein and guide RNA to form ribonucleic protein (RNP) complex.
4. Incubate RNP complex with donor DNA to form pre-initiation complex (PIC).
5. Provide targeted PIC into cells via Electroporation or other means.
6. Allow genome editing/integration to occur for desired time.
7. Ready for screening within hours (Time to detection is cell type and cell cycledependent)
Observe for functional readout (e.g. GFP fluorescence or protein expression) and perform DNA sequencing.