FIGURE

Fig. 2

ID

ZDB-FIG-230118-9

Publication

Yates et al., 2021 - A simple and rapid method for enzymatic synthesis of CRISPR-Cas9 sgRNA libraries

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Fig. 2

Incorporation of the MmeI Recognition Sequence into the Repeat:Anti-Repeat Duplex and Elution with a Strand Displacing Polymerase. (A) Modifications were made to the Repeat:Anti-Repeat Duplex of the sgRNA sequence to insert a single MmeI site that cleaves 18-20 bp upstream of the sgRNA recognition sequence. Modifications are shown in red. The MmeI binding sites are shown in dark gray boxes. The MmeI cut site is represented with a triangle. (B) in vitro Cas9 digestion of a 1000 bp DNA fragment with each sgRNA variant. (C) Injection of zebrafish embryos with Cas9 and the sgRNA variant targeting the gol gene compared to an uninjected control (minimum n per pool = 150). (D) Detachment of the sgRNA scaffold from magnetic beads using Bst 3.0 or Pol1.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nucleic Acids Res.