CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
Type
Blog Post
...to identify the critical NANOG binding sites:
1. Find Enhancers that have NANOG binding sites
2. Design...ChIP-seq experiment (think “GPS for proteins”) for NANOG, an essential transcription factor in embryonic ... just over 13,000 significant binding peaks for NANOG in the human genome. But not all of those 13,000...Design a CRISPR screen to target and disrupt the NANOG enhancers
3. CRISPR out the enhancers from step 2. ... ESCs die or differentiate
Step 3 reveals which NANOG-related genes are critical to stem cell survival...CRISPR in the next section.
In order to denote the NANOG enhancers from step 1 with genome math, we’ll need...shorthand from the field of epigenomics:
H3 – one of NANOG’s associated histone proteins
K4 and K27 – locations...