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Brett T. Staahl
University of California, Berkeley
18Publications
14H-index
2,513Citations
Publications 17
Newest
#1Jun-Jie Liu (University of California, Berkeley)H-Index: 15
#2N. Orlova (California Institute for Quantitative Biosciences)H-Index: 1
Last.Jennifer A. DoudnaH-Index: 98
view all 18 authors...
In this Article, owing to issues with the first 30 nucleotides of the sgRNA, which run in the opposite direction, corrections have been made to the Protein Data Bank (PDB) accessions in the ‘Data availability’ section, and this also affects Figs. 3, 4, Extended Data Fig. 6, Supplementary Table 1 and Supplementary Video 1. The original Article has been corrected online. See the accompanying Amendment for further details.
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#1Jun-Jie Liu (University of California, Berkeley)H-Index: 15
#2N. Orlova (California Institute for Quantitative Biosciences)H-Index: 1
Last.Jennifer A. DoudnaH-Index: 98
view all 18 authors...
The RNA-guided CRISPR-associated (Cas) proteins Cas9 and Cas12a provide adaptive immunity against invading nucleic acids, and function as powerful tools for genome editing in a wide range of organisms. Here we reveal the underlying mechanisms of a third, fundamentally distinct RNA-guided genome-editing platform named CRISPR–CasX, which uses unique structures for programmable double-stranded DNA binding and cleavage. Biochemical and in vivo data demonstrate that CasX is active for Escherichia col...
41 CitationsSource
#1Lucas B. Harrington (University of California, Berkeley)H-Index: 11
#2David Paez-Espino (JGI: Joint Genome Institute)H-Index: 17
Last.Jennifer A. DoudnaH-Index: 98
view all 7 authors...
CRISPR-Cas9 is a powerful technology that has enabled genome editing in a wide range of species. However, the currently developed Cas9 homologs all originate from mesophilic bacteria, making them susceptible to degradation and unsuitable for applications requiring cleavage at elevated temperatures. Here, we show that the Cas9 protein from the thermophilic bacterium Geobacillus stearothermophilus (GeoCas9) catalyzes RNA-guided DNA cleavage at elevated temperatures. GeoCas9 is active at temperatur...
33 CitationsSource
#1Thomas Gaj (University of California, Berkeley)H-Index: 24
#2Brett T. Staahl (University of California, Berkeley)H-Index: 14
Last.David V. Schaffer (HWNI: Helen Wills Neuroscience Institute)H-Index: 61
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Realizing the full potential of genome editing requires the development of efficient and broadly applicable methods for delivering programmable nucleases and donor templates for homology-directed repair (HDR). The RNA-guided Cas9 endonuclease can be introduced into cells as a purified protein in complex with a single guide RNA (sgRNA). Such ribonucleoproteins (RNPs) can facilitate the high-fidelity introduction of single-base substitutions via HDR following co-delivery with a single-stranded DNA...
26 CitationsSource
#1Lucas B. Harrington (University of California, Berkeley)H-Index: 11
#2David Paez-Espino (JGI: Joint Genome Institute)H-Index: 17
Last.Jennifer A. Doudna (University of California, Berkeley)H-Index: 98
view all 7 authors...
CRISPR-Cas9 is a powerful technology that has enabled genome editing in a wide range of species. However, the currently developed Cas9 homologs all originate from mesophilic bacteria, making them susceptible to proteolytic degradation and unsuitable for applications requiring function at elevated temperatures. Here, we show that the Cas9 protein from the thermophilic bacterium Geobacillus stearothermophilus (GeoCas9) catalyzes RNA-guided DNA cleavage over a wide temperature range and has an enha...
5 CitationsSource
#1Brett T. StaahlH-Index: 14
Last.Jennifer A. DoudnaH-Index: 98
view all 10 authors...
Gene editing in the mouse brain is achieved by injection of Cas9 ribonucleoprotein complexes.
99 CitationsSource
#1Benjamin L. OakesH-Index: 9
#2Dana C. NadlerH-Index: 7
Last.David F. SavageH-Index: 21
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The clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated protein Cas9 from Streptococcus pyogenes is an RNA-guided DNA endonuclease with widespread utility for genome modification. However, the structural constraints limiting the engineering of Cas9 have not been determined. Here we experimentally profile Cas9 using randomized insertional mutagenesis and delineate hotspots in the structure capable of tolerating insertions of a PDZ domain without disruption of the enzym...
79 CitationsSource
Cas9, an RNA-guided DNA endonuclease found in clustered regularly interspaced short palindromic repeats (CRISPR) bacterial immune systems, is a versatile tool for genome editing, transcriptional regulation, and cellular imaging applications. Structures of Streptococcus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed protein architecture that undergoes major conformational changes upon guide RNA and DNA binding. To investigate the molecular determinants ...
156 CitationsSource
#1Steven Lin (University of California, Berkeley)H-Index: 25
#2Brett T. Staahl (University of California, Berkeley)H-Index: 14
Last.Jennifer A. DoudnaH-Index: 98
view all 4 authors...
The CRISPR/Cas9 system is a robust genome editing technology that works in human cells, animals and plants based on the RNA-programmed DNA cleaving activity of the Cas9 enzyme. Building on previous work (Jinek et al., 2013), we show here that new genetic information can be introduced site-specifically and with high efficiency by homology-directed repair (HDR) of Cas9-induced site-specific double-strand DNA breaks using timed delivery of Cas9-guide RNA ribonucleoprotein (RNP) complexes. Cas9 RNP-...
477 CitationsSource
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