Andrew Paul May
University of California, Berkeley
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Publication
Featured researches published by Andrew Paul May.
Molecular Cell | 2014
Alexandra E. Briner; Paul Daniel Donohoue; Ahmed A. Gomaa; Kurt Selle; Euan Slorach; Christopher Nye; Rachel E. Haurwitz; Chase L. Beisel; Andrew Paul May; Rodolphe Barrangou
The RNA-guided Cas9 endonuclease specifically targets and cleaves DNA in a sequence-dependent manner and has been widely used for programmable genome editing. Cas9 activity is dependent on interactions with guide RNAs, and evolutionarily divergent Cas9 nucleases have been shown to work orthogonally. However, the molecular basis of selective Cas9:guide-RNA interactions is poorly understood. Here, we identify and characterize six conserved modules within native crRNA:tracrRNA duplexes and single guide RNAs (sgRNAs) that direct Cas9 endonuclease activity. We show the bulge and nexus are necessary for DNA cleavage and demonstrate that the nexus and hairpins are instrumental in defining orthogonality between systems. In contrast, the crRNA:tracrRNA complementary region can be modified or partially removed. Collectively, our results establish guide RNA features that drive DNA targeting by Cas9 and open new design and engineering avenues for CRISPR technologies.
Expert Opinion on Biological Therapy | 2015
Rodolphe Barrangou; Andrew Paul May
The molecular machinery from the prokaryotic clustered regularly interspaced short palindromic repeats (CRISPR)-Cas immune system has broadly been repurposed for genome editing in eukaryotes. In particular, the sequence-specific Cas9 endonuclease can be flexibly harnessed for the genesis of precise double-stranded DNA breaks, using single guide RNAs that are readily programmable. The endogenous DNA repair machinery subsequently generates genome modifications, either by random insertion or deletions using non-homologous end joining (NHEJ), or designed integration of mutations or genetic material using homology-directed repair (HDR) templates. This technology has opened new avenues for the investigation of genetic diseases in general, and for gene therapy applications in particular.
Molecular Cell | 2016
Megan van Overbeek; Daniel Capurso; Matthew Merrill Carter; Matthew S. Thompson; Elizabeth Frias; Carsten Russ; John S. Reece-Hoyes; Christopher Nye; Scott Gradia; Bastien Vidal; Jiashun Zheng; Gregory R. Hoffman; Christopher K. Fuller; Andrew Paul May
Archive | 2014
Andrew Paul May; Rachel E. Haurwitz; Jennifer A. Doudna; James M. Berger; Matthew Merrill Carter; Paul Daniel Donohoue
Trends in Biotechnology | 2017
Paul Daniel Donohoue; Rodolphe Barrangou; Andrew Paul May
Archive | 2015
Peter Cameron; Rachel E. Haurwitz; Andrew Paul May; Christopher Nye; Megan van Overbeek
Archive | 2014
John Van Der Oost; Daniël Christianus Swarts; Andrew Paul May; Rachel E. Haurwitz
Archive | 2017
Paul Daniel Donohoue; Andrew Paul May
Archive | 2017
Matthew Merrill Carter; Overbeek Megan Van; Andrew Paul May
Archive | 2016
Paul Daniel Donohoue; Andrew Paul May; Katharina Friederike Sonja Stengel