Cas9-triggered homologous recombination is a fast and inexpensive way to make essentially any desired change to the genome of C. elegans.
We described this technique in detail in two publications (Dickinson et al. Nature Methods 2013; Dickinson et al. Genetics 2015). We have created this web site as a genome engineering resource for the C. elegans research community. We'll keep it up to date with new protocols, reagent information and answers to questions.
Best of luck with your experiments!
-Dan Dickinson and Bob Goldstein
Best of luck with your experiments!
-Dan Dickinson and Bob Goldstein
New on 8/16/2018: The Invitrogen PureLink HQ miniprep kit, which we recommend for purifying DNA for injection, includes an optional wash step using a 4 M GDN-HCl + 40% isopropanol solution (just before the ethanol wash). We have always included this extra wash step as a precaution, but we never tested whether this was critical. Dave Matus' lab has recently found that it is: omitting the extra wash dramatically reduced their injection efficiency. We have updated our protocols to note that this extra wash step must be included.
New on 5/2/2018: A Micropublication describing our SapTrap cloning kit is now online.
New on 1/19/2018: We have switched to a new online tool, GuideScan (http://guidescan.com) for sgRNA target site selection. We've updated our protocols accordingly.
New on 6/23/2017: We've made a series of constructs that allow use of our SEC selection marker in conjunction with the Jorgensen lab's SapTrap cloning kit. The plasmids are available from Addgene. For more details, see the Constructs and Protocols pages.
New on 5/2/2018: A Micropublication describing our SapTrap cloning kit is now online.
New on 1/19/2018: We have switched to a new online tool, GuideScan (http://guidescan.com) for sgRNA target site selection. We've updated our protocols accordingly.
New on 6/23/2017: We've made a series of constructs that allow use of our SEC selection marker in conjunction with the Jorgensen lab's SapTrap cloning kit. The plasmids are available from Addgene. For more details, see the Constructs and Protocols pages.
