Prof: Your CRISPR editing experiment worked?
Student: CRISPR won’t EDIT, it only CUTS✂️
P: Who edits then?
S: Cell does. But it is unpredictable
P: How can you find out?
S: Using CRISPR-KRISPR
P: What?
S: Yeah, CRISPR-KRISPR, a new method. See paper https://genomebiology.biomedcentral.com/articles/10.1186/s13059-022-02779-8
36s
CRISPRGuru @CRISPRGuru
🧵The CRISPR-KRISPR method was a collaborative work from
@Masato_Ohtsuka
lab. His talented team of bioinformaticians came up with a strategy to simplify Circle-Seq, a popular approach for detecting off target editing events
nature.com
CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
https://www.nature.com/articles/nmeth.4278
We did not find off target cleavages in genetically engineered mouse (GEM) models. It is already well established that off-target cleavages are not common in GEM mice. It is nicely documented in this paper. https://www.nature.com/articles/nmeth.3408
We named our method as CRISPR-KRISPR (CRISPR-Knock-ins and Random Inserts Searching PRotocol)
The method uses very less DNA and it can do much more than just finding out off target cleavages in the genome.