We design, order, and validate CRISPR gRNAs and donor templates needed for knockout, knockin, and deletion modifications for the creation of various animal models as well as for labs to engineer cell lines themselves.
Common types of modifications: KO, deletion, point mutations, small insertions such as floxing and epitope tagging, large insertions
We offer reagent design and validation services for all types of projects, from gene knockouts and targeted mutagenesis to whole locus deletion, targeted insertion, conditional knockout, conditional mutagenesis, and more. Let us know what modifications you would like to make and what target system you would like to make it in, and we will design the experiment strategy, order the reagents, and confirm the activity of these reagents with in-vitro cultured cells before delivery to you or to the mouse engineering core of your choice. This greatly minimizes the risk of expensive technical, manufacturing and design errors in your experiments that can lead to wasted time and effort down the line. Our discounts with synthetic biology reagent companies saves you money on the cost of these materials and the data generated by our validation process provides additional peace of mind that the reagents work.
To submit CRISPR reagent requests, go here, if you are on WashU network. Instructions can be found here. If you are a WU employee working off site, you have to connect to the WashU network via VPN. For non-WU users, please contact us.
Considerations on Mouse Model Creation
GESC works closely with the Transgenic, Knockout and Microinjection Core and Mouse Genetics Core for technology improvements and protocol optimization. In the meantime, we support numerous mouse cores off campus with validated targeting reagents and genotyping services.
- KO and deletion via electroporation of gRNA/Cas9 RNPs: 1-2 sessions
- Point mutations or tagging via electroporation of gRNA/Cas9 RNPs plus one or more single stranded oligodeoxyribonucleotide donors (ssODNs): 1-2 sessions
If animals with a null allele and an allele with desired point mutation are potentially nonviable or infertile, we usually include a block only ssODN to better success rate. Please bring it up during project consultation.