CRISPR/Cas9

In the past targeted gene modification in the mouse genome was only possible through cumbersome gene targeting in embryonic stem cells. This has changed in recent years with the introduction of designer nucleases. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR Associated (Cas) system was originally found as an adaptive immune system of bacteria. Recently, it has been developed for target mutagenesis in the mouse and several other organisms. It became the method of choice for gene modification directly in the oocyte. Cas9 protein is injected together with a guide RNA (gRNA) that determines the specificity, into a mouse zygote. The enzyme will introduce DNA breaks at the location predetermined by the co-injected gRNA. These breaks are repaired by the sometimes faulty non-homologous end joining repair mechanism (NHEJ), leading in a large number of founder animals to random insertions and deletions. When a recombination substrate such as a targeting vector or DNA oligo is co-injected also repair by homologous recombination (HR) can take place, leading to defined gene modifications. 

If you would like to use this new technique for your research, we offer you:

• assistance with the planning
• assistance with the cloning (if necessary)
• the generation of your mice of interest via oocyte injection of gRNA, Cas9 (and targeting vector)

Every Friday we offer a “Targeting Club” at 9:00 am in our seminar rooms (Wagistr. 12, Schlieren or Sternwartstrasse 6, Unispital, Zürich), where you are welcome to discuss your project with us (see also "upcoming dates"). CRISPR/Cas9 projects are a variant of our pronuclear microinjection service, please check there for further information regarding general process description. The prerequisites are at this point project-specific and should be discussed before project start at the "Targeting Club". We are also open to collaborations outside the standard service range.