Degrading DNA with Cascade-Cas3

By Alyssa Shepard

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Graphic showing the workflow of using a pooled AAV CRISPR library in vitro. Step A shows the AAV containing the library with either the gRNA only or gRNA plus Cas9 and infecting cells expressing Cas9 or wild-type. The different guides are represented by different colors in individual cells. Step B shows the results of the positive or negative selection, with only one type of guide (color) being chosen. Step C shows verification using next generation sequencing.
AAV infection of retinal neurons.
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.
Graphic showing different injection routes in mice; see text for details

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