CRISPR/Cas9-based genome editing at the Transgenesis Core Facility
We have experience and expertise in using high efficiency CRISPR/Cas9 approaches to generate transgenic mice with:
- gene knock-outs
- conditional knock-outs
- variant allele knock-ins
- transgene knock-ins
Our in house protocols are based on the pioneering work of Channabasavaiah Gurumurthy and Masato Ohtsuka who developed the Easi-CRISPR method of using long ssDNA donors and CRISPR ribonucleoproteins to generate transgenic mice with high efficiencies [1, 2].
We provide advice and assistance to researchers on:
- the development of CRISPR/Cas9-based genome editing strategies
- the selection of suitable gRNAs
- the design of ssDNA repair templates
- strategies for the genotyping and genetic characterization of founder mice
- the establishment and maintenance of transgenic mouse lines
We are continually working to improve on existing methods and to develop new approaches which will allow us to design more efficient and successful CRISPR/Cas9-based gene editing strategies and provide us with the tools to tackle more challenging gene editing projects.
We actively encourage all researchers who are interested in using (or who currently use) CRISPR/Cas9 based genome editing approaches in mice to come and discuss their projects with us, because each gene-editing project is different and presents its own unique challenges. The sharing of knowledge and experience is essential to finding solutions to the most challenging of gene editing projects as valuable information is gained not only from experimental approaches which were successful, but also from experimental designs that were unsuccessful. If you have an idea of an experiment that you would like to try come and talk to us and we will see if we can help you make it a reality.
1. Quadros, R.M., et al., Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins. Genome Biol, 2017. 18(1): p. 92.
2. Miura, H., et al., Easi-CRISPR for creating knock-in and conditional knockout mouse models using long ssDNA donors. Nat Protoc, 2018. 13(1): p. 195-215.