Are you looking for a simple means to boost the efficiency of gene editing and also to reduce the off-target? Yes, it does exist – in one word: 4-hydroxy Tamoxifen. That’s the HIT-CAS9.

Hybrid drug inducible CRISPR/Cas9 technology, HIT-Cas9

Zhao et al revealed tight and effective drug control of the CRISPR Cas9 based on 4-Hydroxytamoxifen. doi: 10.1016/j.omtn.2018.08.022

They built a fusion between a mutated human estrogen receptor (ERT2) and the Cas9 endonucelase, which makes the CRISPR Cas9 dependent on 4-hydroxytamoxifen.

Optimal knock-down can be done by a shRNA sequence that depends on the gene expression level and the cell type. Usually, 4 shRNA should be tested to find the one inducing a minimum of 70% increase of the targeted mRNA pool. Genecopoeia has developped shRNA expression vectors with improved performances.

CRISPR-CAS9 is a powerful technology for gene editing. It allows targeted modifications into the genome of the cell lines, IPSCs and ESCs. Despite the principle being pretty simple, in practice its use requires some expertise, and it can even turn into a time consuming adventure… That’s why the best way to benefit from it is most certainly through affordable services.

RNAi offer >70% knock-down of the gene expression, and thanks to CRISPR-CAS9 gene editing, it’s possible make a complete knock-out. The TUNR system offers the possibility to obtain intermediates with high, medium and low reduction of the gene expression and with the same quality of targeting as for knock-out. Let’s take a closer look at the TUNR technique.