Protocols & Reagents

Protocols

We have deposited many of the lab’s detailed protocols on Protocols.IO at http://www.protocols.io/groups/cornlab.

Some of the most popular protocols are

• Cas9 expression and purification
• In vitro transcription of guide RNAs
• Nucleofection of Cas9 RNP for high efficiency genome editing
• T7 Endonuclease (T7E1) assay
• Reamplification of CRISPR libraries

Example T7E1 digestion (treated vs untreated) for thirteen genes that were targeted with the Cas9 RNP. These results are representative for a larger set of 200 genes targeted in this experiment.

Reagents and Cell Lines

We make most of the lab’s published (and some unpublished) reagents available on Addgene at https://www.addgene.org/Jacob_Corn/. Email us if you read about a reagent in one of our papers and can’t find it on Addgene (e.g. HDR donor oligos, in vitro transcribed sgRNAs).

All of our CRISPR stable cell lines (e.g. for genome-wide screening) are available through the Berkeley Cell Culture facility. Below are a few examples of CRISPRi/a lines we have deposited through the facility. Check the link above for an up-to-date list of lines.

• Jurkat dCas9-VPR (CRISPRa)
• Jurkat dCas9-KRAB (CRISPRi)
• HCT116 dCas9-KRAB (CRISPRi)
• 3T3-L1 dCas9-KRAB (CRISPRi)

Contact Allison Killilea at ankillilea@berkeley.edu for more information on Corn lab CRISPR lines. Cell lines described in papers but not distributed by the Berkeley Cell Culture facility are available by direct request.