GeneCopoeia's Safe Harbor Knock-in ORF clones are designed for transfering the ORF of your gene of interest from a donor plasmid into the safe harbor sites in the human or mouse genomes via CRISPR or TALEN-mediated homologous recombination (HR; Ffigure 1). Over 20,000 sequence-verified human ORFs and over 15,000 mouse ORFs are available. Safe Harbor Knock-in ORF clones are designed for use with GeneCopoeia's Safe Harbor Gene Knockin Kits.
Figure 1. Genome-editing-tool-mediated transgene integration at the safe harbor sites
Figure 2. Maps of (A) Human safe harbor AAVS1 knock-in ORF clone and (B) Mouse safe harbor ROSA26 knock-in ORF clone
Figure 3: Human genome safe harbor AAVS1 gene targeting of OCT4 and SOX2.
(A) OCT4 (Cat#: DC-Z0092-SH01) or SOX2(Cat#: DC-T2547-SH01) donor plasmids were co-transfected with AAVS1 TALEN Pair into HEK293T cells in a 6-well pate. Cells were sub-cultrured 48 hr post-transefection and selected against puromycin (1µg/ml) for 2 weeks. The expression of CopGFP was checked under a microscope (Nikon Eclipse Ti ) 48h post-transfection or after 2 weeks of drug selection. Few colonies left in cells transfected with Donor plasmid alone after puromycin selection (data not shown).
(B) Western blot analysis of proteins from HEK293T cells stably integrated with SOX2(Cat#: DC-T2547-SH01) or OCT4 (Cat#: DC-Z0092-SH01) in AAVS1 site, with cells alone as negative control where endogenous Sox2 or OCT4 protein levels were too low to be detected in the same blot.
Human and Mouse Safe Harbor Knockin Kits
Human AAVS1 and mouse ROSA26 safe harbor knockin kits – CRISPR- or TALEN-mediated gene targeting kits designed to specifically transfer your gene of interest, selection marker or other genetic element into the human AAVS1 or mouse safe harbor sites. More than 35,000 AAVS1- and ROSA26-compatible ORF donor clones are also available. The safe integration ensures transcription-competency of the transgenes and presents no known adverse effects on fitness.
Cas9 Stable Cell Lines
Genome-CRISP™ CRISPR-Cas9 stable cell lines - Pre-made human cell lines H1299, HEK293T, HeLa, and A549 with the CRISPR Cas9 nuclease stably integrated into the human AAVS1 “Safe Harbor” site.
CRISPR and TALEN Products and Services
Genome-CRISP™ 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.
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.