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CRISPR Plasmids: Cut


Fully functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often lead to frameshifts, creating loss of function alleles.

To introduce specific genomic changes, researchers use ssDNA or dsDNA repair templates with 1. homology to the DNA flanking the DSB and 2. a specific edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs at a much lower efficiency than NHEJ.

CRISPR Cut illustration depicts complex formation, target binding, and double-strand break formation (target cleavage). Repair of the break by nonhomologus end-joining joins the ends together with minimal processing and results in insertions and deletions, which can lead to frameshift mutations. If a repair template is present with homologous arms and a desired genomic edit close to the PAM site, homology-dependent repair can copy template sequence into the break site resulting in a precise edit.


Want more information on the wide variety of Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations, and more, created and maintained by the Doudna Lab.

Browse, sort, or search the tables below for CRISPR plasmids designed to introduce a DSB.
Plasmids are available for expression in mammalian systems, bacteria, Drosophila, plants, C. elegans, yeast, zebrafish, and Xenopus.


Mammalian

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Bacteria

Plasmid Gene/Insert Promoter PI Publication

Drosophila

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Plant

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

C. elegans

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Yeast

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Zebrafish

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Xenopus

Plasmid Gene/Insert Promoter Selectable Marker PI Publication

Parasites

Plasmid Gene/Insert Promoter Selectable Marker PI Publication


Do you have suggestions for other plasmids that should be added to this list?

Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!