The Addgene Blog

Master plasmid fundamentals, CRISPR techniques, AAV serotype selection, and antibody applications. Written by scientists, for scientists.

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A cartoon representation of a bacterium containing six different-colored plasmids. A zoomed-in view of one plasmid shows the synthetic plasmid ORI producing an RNA repressor, which represses replication of the plasmid.
A heatmap showing on-target activity of SpCas9 variants against various targets, with WT-like cleavage indicated by green, no cleavage indicated by red, and intermediate cleavages in middle shades of yellow-green, yellow, and orange. The variants are ranked by activity and the targets are ranked by cleavability, with the result that the top-left corner (high activity, high cleavability) is green to show high cleavage, the bottom right corner (low activity, low cleavability) is red to show no cleavage, and an approximate staircase pattern divides the two colors. In each row, a miniaturized graph of off-target cleavage, normalized from 0 to 1, is shown. In general, these graphs show declining off-target cleavage among SpCas9 variants of lower activity. In each row, representing a particular target, one cell of the heatmap has been outlined in black to emphasize it as belonging to the target-matched variant. Each target-matched cell is the last in its row to show moderate yellow-green color rather than an orange color, indicating some on-target cleavage activity. Each target-matched variant has very low off-target cleavage.
A protein structure of ABE8e is shown, superimposed with cartoon representations of sgRNA and target DNA. Two TadA domains are highlighted. The catalytic domain contacts multiple bases of the non-target DNA strand, while the docking domain primarily contacts the other protein domains.
A photo of three people holding and touching a tray of small plants that are either bright green or dark purple.

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