Transgene Knockin via CRISPR at AAVS1 and ROSA26 Loci

Transgene expression via random integration into the genome is subjected to position effects and silencing. In addition, random gene insertion might interrupt or activate the neighboring genes. Genomic safe harbor sites are transcriptionally active, therefore allowing robust and stable gene expression. Furthermore, a transgene insertion at genomic safe harbors does not have adverse effect on the host cell genome. CRISPR technology can be utilized to do targeted gene insertion at these genomic loci. For Human cells, AAVS1 has been accepted as a high gene expression and a safe genomic location; for Mouse cells, ROSA26 is proved to be a genomic safe harbor locus.

This system needs two components:

AAVS1 Knockin Validation Data

1. AAVS1 gRNA vector,All-in-one CRISPR vector containing Cas9 expression cassette and specific gRNA targeting AAVS1 locus
2. AAVS1 Donor vector,donor vector for transgene integration, containing the AAVS1 homologous arms (Gene of interest needs to be cloned)

OriGene offers a prevalidated AAVS1 gRNA and many AAVS1 donor vectors, which contains different combination of drug selection marker and promoter. In addition to the individual gRNA and donor vector, the pre-assembled AAVS1 vector kit is also available, including not only AAVS1 specific gRNA and donor vector, but also a non-specific scramble control. You can also get our predesigned positive donor control for your experiment or request a custom AAVS1 donor plasmid.

This system needs two components:

1. pCas-Guide-ROSA26, All-in-one CRISPR gRNA vector for ROSA26 targeting
2. pAAVS1-Puro-DNR, donor vector for transgene integration, containing ROSA26 homologous arms (Gene of interest needs to be cloned)

ROSA26 Knockin validation data