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CRISPR Guide Update

By Alyssa Shepard

Alyssa Shepard October 10, 2024

Addgene is proud to present our updated CRISPR Guide!

The CRISPR Guide helps scientists understand how to use CRISPR plasmids in the lab and the vast range of possibilities at their disposal. With this update, we have added new sections to share and describe some incredible new CRISPR technologies developed and widely adopted in the past few years, as well as refreshing the existing content.

Access the CRISPR Guide here!

Highlights include a much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements and enhanced editing, as well as alternatives to Cas9. All of this is accompanied by new or refreshed figures made using BioRender.com.

CRISPR prime editing schematic. The parts of the prime editor and pegRNA are indicated. The prime editor consists of a Cas9n H840A nickase fused to a reverse transcriptase (RT), while the pegRNA includes spacer, scaffold, RT template, and primer binding site (PBS) sequences. The desired edit is part of the RT template. In the first step, these components form a complex, bind target DNA, and nick the Cas9 non-target strand. Next, the primer sequence binds the freed non-target strand, and the RT extends it using the RT template, incorporating the edit into target DNA. Finally, the DNA is freed from the prime editing complex, and cellular endonucleases and mismatch repair resolve the heteroduplex.
Figure 1: PRIME editing

Please check out the updated guide for a great introduction to what CRISPR technologies have to offer. Happy CRISPR-ing!

More resources on the Addgene blog

PRIME Editing

CRISPR 101: Cas9 vs the Other Cas(s)

Progress Towards a PAM-free CRISPR

Topics: CRISPR, Addgene News

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