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CRISPR Plasmids: Nick


Cas nickase mutants introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild-type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed repair (HDR).

A schematic of Cas9n used to create DSB. gRNA and gRNA2 are added to Cas9n. An arrow indicates complex formation and target binding. Two Cas9n and gRNAs are bound to a length of double-stranded DNA, one to Target 1 and PAM 1, one to Target 2 and PAM 2, which is ~12 bases away from PAM 1. An arrow indicates Target cleavage and DSB formation. The double-stranded DNA, no longer bound by Cas9n complexes, shows three bases missing on the 5' strand on Target 1, next to the PAM 1 site, and 3 bases missing on the 3' strand on Target 2, next to the PAM 2 site. An arrow indicates DSB repaired via NHEJ or HDR.
Figure 1: Overview of Cas9 nickase cleavage and repair.

Browse, sort, or search the tables below for CRISPR nickase plasmids. To learn more about CRISPR nickases and other CRISPR topics, read our CRISPR Guide.

Mammalian

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Bacteria

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Drosophila

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Plant

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Yeast

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CRISPR Resources

Addgene has a large selection of CRISPR plasmids and resources. Find more CRISPR functions along with plasmids categorized by organism by visiting our CRISPR plasmids page. Find a comprehensive list of CRISPR resources by visiting our CRISPR reference page.


Content last reviewed: 17 October 2025

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