The enChIP Kit (engineered DNA-binding molecule-mediated chromatin immunoprecipitation) utilizes the CRISPR/Cas system to target specific genomic regions for investigation of associated DNA-binding interactions.
A guide RNA (gRNA) containing 20 nucleotides complementary to the desired genomic region is expressed in combination with a tagged, enzymatically inactive Cas9 protein (dCas9). The gRNA directs the dCas9 protein to the target sequence and an RNA-DNA heteroduplex is formed. Chromatin immunoprecipitation is then performed on the cells using an antibody directed against the AM-tagged dCas9 protein to enrich for genomic sequences bound by the gRNA/dCas9 complex.
By using the enChIP Kit, off-target gRNA binding sites can be identified. This provides valuable information regarding the quality of gRNA design prior to use in genome editing experiments. Additionally, the enChIP Kit can be used to study cis– and trans-interacting chromosomal looping events.
- Identify off-target binding events for each gRNA sequence
- Evaluate cis– and trans-interacting chromosomal looping events
- AM-tag design maximizes enChIP enrichment efficiency
- Works well with open chromatin regions to detect promoter, enhancer and insulator elements