AAVS1 Safe Harbour Integration-TALEN tools and services
Genome-TALER™ human AAVS1 safe harbor gene targeting kit is designed to specifically transfer your gene of interest, selection marker or other genetic element from a donor plasmid into the AAVS1 safe harbor site in human chromosome 19 via TALEN-mediated homologous recombination
| Supplier |
|---|
| LabOmics-GeneCopoeia |
| Product |
| Genome-TALER™ Human AAVS1 Safe Harbor Gene Targeting Kit |
Features
- Safe Harbour Chromosomal Integration: designed to specifically transfer your gene of interest, selection marker or other genetic element from a donor plasmid into the AAVS1 safe harbor site in human chromosome 19
- TAL Technology: via very specific TALEN-mediated homologous recombination (HR)
- Known copy number Known copy number of the transgene ensures predictable expression level and simplifies phenotype interpretation
- Compatible knock-in ORFs: Over 18,000 sequence-verified human ORFs are compatible for transgene donor DNA design
The modification of the human genome by insertion of genes of interest and other genetic elements in unique site(s) of chromosome(s) is of great value for cell engineering. The genetically modified cells are valuable for therapeutic research, gene function study as well as lineage tracking and analysis. All these applications depend on the reliable and predictable function of the transgene without perturbing any endogenous gene and/or other regulation element. Random integration of the transgene, on the contrary, can present a threat of unpredicted insertion or mutagenesis.
The new approach recently developed is to deliver the transgene to a predetermined and safe site in a genome. AAVS1 (also known as PPP1R2C locus) in human chromosome 19, is a well-validated “safe harbor” to host the DNA fragment with expected function. It has an open chromatin structure and is transcription-competent. Most importantly, there is no known adverse effect on the cell resulting from the inserted DNA fragment of interest.
Genome-TALER™ human AAVS1 safe harbor gene targeting kit is designed to specifically transfer your gene of interest, selection marker or other genetic element from a donor plasmid into the AAVS1 safe harbor site in human chromosome 19 via TALEN-mediated homologous recombination (HR). HR is a natural DNA repair mechanism that occurs in response to DNA double-strand breaks (DSBs). The DSBs here are created by AAVS1-specific TALENs (figure 1).
Figure 1. Illustration of TALEN-mediated transgene integration at the safe harbor AAVS1 site of human genome
The new approach recently developed is to deliver the transgene to a predetermined and safe site in a genome. AAVS1 (also known as PPP1R2C locus) in human chromosome 19, is a well-validated “safe harbor” to host the DNA fragment with expected function. It has an open chromatin structure and is transcription-competent. Most importantly, there is no known adverse effect on the cell resulting from the inserted DNA fragment of interest.
Genome-TALER™ human AAVS1 safe harbor gene targeting kit is designed to specifically transfer your gene of interest, selection marker or other genetic element from a donor plasmid into the AAVS1 safe harbor site in human chromosome 19 via TALEN-mediated homologous recombination (HR). HR is a natural DNA repair mechanism that occurs in response to DNA double-strand breaks (DSBs). The DSBs here are created by AAVS1-specific TALENs (figure 1).
Figure 1. Illustration of TALEN-mediated transgene integration at the safe harbor AAVS1 site of human genome
This kit is designed to specifically transfer your gene of interest, selection marker or other genetic element into the AAVS1 safe harbor site in human chromosome 19 via TALEN-mediated homologous recombination. The kit includes one pair of AAVS1 TALENs, one donor cloning vector or one donor clone*, one positive control clone and one knock-in PCR verification kit.
Figure 2. Safe harbor gene targeting kit. (A) TALEN and donor DNA plasmids (B) Knock-in verification PCR primers
* AAVS1 donor clones can be custom-made and included in the kit instead of the empty AAVS1 donor cloning vector.
Figure 3. Human genome safe harbor AAVS1 gene targeting AAVS1 donor control plasmid pAAVS1D-RFP-copGFPpuro (800 ng) was co-transfected with AAVS1 TALEN pair(600 ng for each) or control TALEN pair into HEK293T cells in a 6-well pate. (B) 48 hr post-transfection, the cells were split 1:10 into a new 6-well pate and selected against 1.0 µg/ml of puromycin. The images were taken after two weeks of selection. The cells transfected with pAAVS1D-RFP-copGFPpuro + control TALEN were completely killed by puromycin selection (data not shown). (D) PCR primers designed to amplify the HR junction were used to verify the successful integration.
| Cat. No | Description | Includes | Price |
|---|---|---|---|
| SH-AVS-K100 | AAVS1 safe harbor gene targeting kit | AAVS1 TALEN pair (TN-AAVS1) AAVS1 donor cloning vector (DC-DON-SH01) AAVS1 positive control donor (DC-RFP-SH01) knock-in verification primer pairs (HQPAVSHR) | € 1810 |
| TN-AAVS1 | AAVS1 TALEN pair | AAVS1 left TALEN AAVS1 right TALEN | N/A |
| DC-DON-SH01 | AAVS1 donor cloning vector | Cloning vector for donor DNA construction Includes MCS, eGFP reporter, and Puromycin marker | € 580 |
| DC-RFP-SH01 | AAVS1 positive control donor | RFP donor clone for positive control | € 580 |
| HQPAVSHR | AAVS1 knock-in verification PCR primer pairs | Primer pair for 5’ HR junction Primer pair for 3’ HR junction | N/A |
| Variable | AAVS1 donor clone custom services | Custom-made AAVS1 donor clone containing your GOI or other genetic element for targeted AAVS1 integration | Custom services |
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