Introduction
TALEN kits The kits are 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 Genome-TALER™ human AAVS1 safe harbor gene knock-in kit includes one pair of AAVS1 TALENs, one donor cloning vector or one donor clone*, one positive control clone and two pairs of knock-in PCR verification primers.
Figure 1. Genome-TALER™ human AAVS1 safe harbor gene knock-in kit. (A) AAVS1 TALENs and donor DNA plasmids (B) AAVS1 knock-in verification PCR primers
* DC-DON-SH01 only comes with SH-AVS-K100 kit. Genome-TALER™ AAVS1 ORF knock-in clones can be and ordered separately. Other AAVS1 donor clones can also be custom-made by us.
Acknowledgement: Design of the AAVS1 left TALEN, AAVS1 right TALEN, and AAVS1 donor control vectors was performed by Dr. Jizhong Zou of the NIH Center for Regenerative Medicine, a Common Fund initiative of the U.S. National Institutes of Health.
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| TN-AAVS1-L | AAVS1 left TALEN | Specifically cleave AAVS1 with target site TN-AAVS1-R to create DSB | 1 |
| TN-AAVS1-R | AAVS1 right TALEN | Specifically cleave AAVS1 with target site TN-AAVS1-L ​to create DSB | 1 |
| DC-DON-SH01* | AAVS1 donor cloning vector |
For cloning GOI or containing GOI –Two AAVS1 flanking arms for HR -CMV promoter -GFP/Puro marker for selection |
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| DC-RFP-SH01 | AAVS1 positive control donor clone | RFP instead of GOI for positive control purpose | 1 |
| HQPAVSHR-5 | 5′ knock-in verification PCR primer pair | PCR primer pairs for 5’ HR junctions to verify successful integration | 1 |
| HQPAVSHR-3 | 3′ knock-in verification PCR primer pair | PCR primer pairs for 3’ HR junctions to verify successful integration | 1 |
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Figure 2. Human genome safe harbor AAVS1 gene targeting (A) AAVS1 RFP control plasmid DC-RFP-SH01 (800 ng) was co-transfected with AAVS1 TALEN pair(600 ng for each) or control TALEN pair into HEK293T cells in a 6-well plate. (B) 48 hr post-transfection, the cells were split 1:10 into a new 6-well plate and selected against 1.0 µg/ml of puromycin. The images were taken after two weeks of selection. Few colonies left in the wells transfected with AAVS1 RFP control + control TALEN. (C) (D) PCR primers designed to amplify the HR junction were used to verify the specific and successful integration. |
CRISPR kits The kits are 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 CRISPR-Cas9-mediated homologous recombination. The GeneHero™ human AAVS1 safe harbor gene knock-in kit includes one AAVS1 sgRNA/Cas9 expression clone, one donor cloning vector or one donor clone*, one positive control clone and one knock-in PCR verification kit.
Figure 3. GeneHero™ human AAVS1 safe harbor gene knock-in kit. (A) AAVS1 sgRNA/Cas9 expression clone and donor DNA plasmids (B) AAVS1 knock-in verification PCR primers
* DC-DON-SH01 only comes with SH-AVS-K200 kit. Genome-TALER™ AAVS1 ORF knock-in clones can be and ordered separately. Other AAVS1 donor clones can also be custom-made by us.
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| Cat. No. | Components | Description | Qty |
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| HCP-AAVS1-CG02 | AAVS1 sgRNA/Cas9 expression clone | Specifically cleave AAVS1 target site to create DSB | 1 |
| DC-DON-SH01* | AAVS1 donor cloning vector |
For cloning GOI or containing GOI –Two AAVS1 flanking arms for HR -CMV promoter -GFP/Puro marker for selection |
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| DC-RFP-SH01 | AAVS1 positive control donor clone | RFP instead of GOI for positive control purpose | 1 |
| HQPAVSHR-5 | 5′ knock-in verification PCR primer pair | PCR primer pairs for 5’ HR junctions to verify successful integration | 1 |
| HQPAVSHR-3 | 3′ knock-in verification PCR primer pair | PCR primer pairs for 3’ HR junctions to verify successful integration | 1 |
Knock-in clones Human AAVS1 safe harbor knock-in ORF clones More than 20,000 sequence-validated human AAVS1 safe harbor knock-in ORF clones are available. These clones are constructed for specifically integrating the ORF of your interest from a donor plasmid into the AAVS1 safe harbor site in human chromosome 19 by using Genome-TALER™ or GeneHero™ human AAVS1 safe harbor gene knock-in kits.
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Figure 4. Map of human AAVS1 safe harbor knock-in ORF clones
Human AAVS1 Safe Harbor knock-in CRISPR-Cas9 clones
| Buy | Catalog # | Product | Application | Selection marker | Promoter | Cas9 integration site | Delivery format | Price |
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| SH300 | Cas9 AAVS1 knockin donor clone | To knock in Cas9 nuclease to human AAVS1 Safe Harbor site using CRISPR or TALEN | Puro | CBh | AAVS1 | Bacterial stock or transfection-ready DNA | $495 | |
| SH301 | Cas9 AAVS1 knockin donor clone | Hygro | CBh | AAVS1 | $495 | |||
| SH302 | Cas9 AAVS1 knockin donor clone | Neo | CBh | AAVS1 | $495 |
| User Manual |
Knock-in clone custom services also available
GeneCopoeia also offers AAVS1 donor clone custom services to construct knock-in donor clones containing mutated genes or other genetic elements for targeted AAVS1 integration. To get a quote, please click here. Questions? Please contact us at inquiry@genecopoeia.com or 1-301-762-0888.
Services
Questions? Please contact us at inquiry@genecopoeia.com or 1-301-762-0888.
AAVS1 donor clone custom services
GeneCopoeia also offers AAVS1 donor clone custom services to construct knock-in donor clones containing mutated genes or other genetic elements for targeted AAVS1 integration. To get a quote, please click here. Questions? Please contact us at inquiry@genecopoeia.com or 1-301-762-0888.AAVS1 stable cell line custom services
| Services | Description | Application | ||
|---|---|---|---|---|
| Stable cell line services | Monoclonal colony | Monoclonal stable cell line with TALEN- or CRISPR-Cas9-mediated safe harbor transgene. | human AAVS1 | |
| Cell bank | Create cell bank of monoclonal stable cell line with TALEN or CRISPR-Cas9-mediated safe harbor transgene. | human AAVS1 | ||
Related Products
Mouse ROSA26 safe harbor kit
Genome-TALER™ or Genome-CRISP™ mouse ROSA26 safe harbor gene knoc-in kits and clones – TALEN- or CRISPR-mediated gene knock-in kits designed to specifically transfer your gene of interest, selection marker or other genetic element into the ROSA26 safe harbor site in mouse chromosome 6. More than 20,000 ROSA26 compatible ORF donor clones are also available. The safe integration ensures transcription-competency of the transgenes and presents no known adverse effect.
TALE and CRISPR-Cas9 products and services Genome-TALER™ TALEN and TALE-TF custom services – GeneCopoeia offers several levels of affordable custom services for designing, creating and validating TALENs, TALE-TFs and other TAL effectors-based targeted genomic modification tools. GeneHero™ CRISPR-Cas9 products and services – GeneCopoeia offers CRISPR-Cas9 sgRNA design and cloning services, Cas9 expression clones and other related products and services for simple and fast targeted genomic editing.
Cas9 stable cell lines GeneHero™ CRISPR-Cas9 stable cell lines – GeneCopoeia offers stable cell lines constitutively expressing the CRISPR Cas9 nuclease for a convenient means to carry out CRISPR genome editing applications with high efficiency, especially for the highthoroughput sgRNA library screening.
Resources
Product Datasheets (Sales Order # is required). Application Notes
GeneCopoeia genome editing tools for safe harbor integration in mice and humans- FAQs FAQs about AAVS1 safe harbor gene targeting kits and clones and other genome editing products
- Webinar Watch recorded webinar: Solutions for genome editing: TALEN & CRISPR-Cas9, and Safe Harbor Knock-in ORF clones Download webinar PPT slides
- Documents User manual: Genome-TALER™ Human AAVS1 safe harbor gene knock-in kit user manual
- MSDS: Genome-TALER™ Human AAVS1 safe harbor gene knock-in kit
- Warranty statement: Genome-TALER™ AAVS1 knock-in ORF donor clones Cancellation, Quality Assurance, Limited Use License and Warranty