The CRISPR/Cas9 system is a recent and rapidly evolving technique for genetic manipulation. With time we envisage that it will become an essential part of our existing and future projects.
The clustered regularly interspaced short palindromic repeat (CRISPR) construct is used in combination with guide RNA (gRNA), which contains a protospacer complementary to the target sequence, and either the CRISPR-associated protein Cas9, which cleaves the target site, or Cas9D10A, a nickase. Together, these form a complex that can locate, bind to and cleave or nick a specific target gene. The cell’s own repair mechanisms then replace this section of DNA with an oligonucleotide containing the desired mutation.
From start to finish, this process takes approximately eight weeks to complete, whereas our current system requires a lengthy breeding programme and the whole process can last for months. This means that, by using the CRISPR/Cas9 system, we will be able to produce mutant mice considerably cheaper, faster and more efficiently. It will also reduce the numbers of mice we require to establish a mutant mouse line.
While there are still some limitations to CRISPR/Cas9, it offers huge benefits and the rate of change is fast, with improvements and adaptations being developed by our own researchers and other external groups. We intend to make extensive use of the promise this method shows, and have begun to incorporate it into our current pipelines. We are therefore currently pursuing initial exploratory research in order to determine how best to use this new technique and establish it within our existing framework.