CASTing Off for New Shores in Human Genome Editing

By Emily P. Bentley

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Diagram of the Cascade complex with gRNA bound to target DNA, after recruitment of Cas3. Cas3 is about to nick the non-target strand.
A cartoon of a prime editor with two different edit sequences. The DNA sequences are shown with one strand edited and a 5′ DNA flap, before heteroduplex resolution and DNA repair.  The first edit has an unchanged PAM. This DNA is shown connected to the prime editor by a two-way arrow, indicating that the editor can re-bind. Re-nicking is represented by scissors and would remove the newly edited DNA.  The second edit has an altered PAM. A one-way arrow leads from the prime editor to this edit, indicating that the changed PAM prevents the editor from re-binding.
A cartoon overlayed on several crystal structures showing the parts of the prime editor: the Cas9 nickase domain, the reverse transcriptase domain, and the pegRNA.
screenshot of various PRIDICT webpages
schematic representation of the PASTE system
Using CRISPR in C. elegans to knock-in FLAG tag.
Multiplexed insertion is possible using a single plasmid that contains a multi-spacer CRISPR array. This encodes separate gRNAs that target different locations in the genome.

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