Disruptive non-disruptive applications of CRISPR/Cas9

- Using Cas9 as a generic DNA binding platform offers novel ways to study the genome.
- Specific bases can be edited using Cas9.
- Transcription or epigenetic imprinting of genetic loci can be reprogrammed using Cas9.
- Genomic loci can be visualized and cellular lineage trees can be derived using Cas9.
- Cas9-based genomic recording and circuits allow to store and process data in cells.

The bacterial type II Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR Associated (Cas) systems, and in particular Streptococcus pyogenes CRISPR-Cas9, have been broadly applied to edit the genome of bacterial and eukaryotic cells. Cas9, which is an RNA-guided programmable nuclease, is a powerful tool for disrupting protein-coding genes. Cas9 cleaves target sites to generate a double-strand break (DSB) that is repaired via an error-prone repair process, leading to insertion/deletion mutations and gene knockouts. However, Cas9 can also be used to modulate genome function without gene disruption, enabling base editing, transcriptional and epigenetic reprogramming, genome imaging, cellular barcoding, genetic recording, and genetic computation.

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