ssDNA and the Argonautes: The Quest for the Next Golden Editor
Argonaute is a DNA-guided DNA cursor that represents an exciting new addition to the genome engineer's toolbox.4
To cite this article: Martínez-Gálvez Gabriel, Ata Hirotaka, Campbell Jarryd M., and Ekker Stephen C.. Human Gene Therapy. June 2016, 27(6): 419-422. doi:10.1089/hum.2016.071.
Published in Volume: 27 Issue 6: June 20, 2016 Online Ahead of Editing: May 27, 2016Gabriel Martínez-Gálvez,1Hirotaka Ata,2Jarryd M. Campbell,2 and Stephen C. Ekker2,*
1Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota.
2Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota.
*Correspondence: Dr. Stephen C. Ekker, Department of Biochemistry/Molecular Biology, Mayo Clinic, 200 1st Street SW, Guggenheim 1342C, Rochester, MN 55905. E-mail:ekker.stephen@mayo.edu
Received for publication May 25, 2016 accepted after revision May 26, 2016
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The RNA-guided endonuclease Cas9 has made genomeediting a widely accessible technique. Similar to Cas9, endonucleases from theArgonaute protein family also use oligonucleotides as guides to degrade invasive genomes. Here we report that the NatronobacteriumgregoryiArgonaute (NgAgo) is a DNA-guided endonuclease suitable for genomeediting in human cells. NgAgo binds 5' phosphorylated single-stranded guide DNA (gDNA) of ～24 nucleotides, efficiently creates site-specific DNA double-strand breaks when loaded with the gDNA. The NgAgo-gDNA system does not require a protospacer-adjacent motif (PAM), as does Cas9, and preliminary characterization suggests a low tolerance to guide-target mismatches and high efficiency in editing (G+C)-rich genomic targets.
