Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA
| العنوان: | Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA |
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| المؤلفون: | Osamu Nureki, Yinqing Li, Feng Zhang, Ryuichiro Ishitani, Takanori Nakane, Kira S. Makarova, Ian Slaymaker, Hisato Hirano, Hiroshi Nishimasu, Bernd Zetsche, Iana Fedorova, Eugene V. Koonin, Takashi Yamano |
| المساهمون: | Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Biology, Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research at MIT, Ran, Fei, Hsu, Patrick, Konermann, Silvana M, Shehata, Soraya I., Zhang, Feng |
| المصدر: | PMC Cell, vol 156, iss 5 |
| بيانات النشر: | Elsevier BV, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Models, Molecular, CRISPR/Cpf1, CRISPR-Associated Proteins, Crystallography, X-Ray, Medical and Health Sciences, chemistry.chemical_compound, Endonuclease, 0302 clinical medicine, Models, Guide RNA, Kinetoplastida, Genetics, 0303 health sciences, Crystallography, biology, Bacterial, Biological Sciences, 3. Good health, RNA, Bacterial, Protospacer adjacent motif, Infectious Diseases, RNA, Guide, Kinetoplastida, Heteroduplex, DNA, Bacterial, Protein Structure, Streptococcus pyogenes, 1.1 Normal biological development and functioning, Molecular Sequence Data, Computational biology, Article, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Underpinning research, Amino Acid Sequence, 030304 developmental biology, Nuclease, Bacteria, Biochemistry, Genetics and Molecular Biology(all), Cas9, Human Genome, Molecular, DNA, Endonucleases, Protein Structure, Tertiary, Emerging Infectious Diseases, chemistry, X-Ray, biology.protein, RNA, Sequence Alignment, Tertiary, Guide, 030217 neurology & neurosurgery, Developmental Biology |
| الوصف: | The CRISPR-associated endonuclease Cas9 can be targeted to specific genomic loci by single guide RNAs (sgRNAs). Here, we report the crystal structure of Streptococcus pyogenes Cas9 in complex with sgRNA and its target DNA at 2.5 Å resolution. The structure revealed a bilobed architecture composed of target recognition and nuclease lobes, accommodating the sgRNA:DNA heteroduplex in a positively charged groove at their interface. Whereas the recognition lobe is essential for binding sgRNA and DNA, the nuclease lobe contains the HNH and RuvC nuclease domains, which are properly positioned for cleavage of the complementary and noncomplementary strands of the target DNA, respectively. The nuclease lobe also contains a carboxyl-terminal domain responsible for the interaction with the protospacer adjacent motif (PAM). This high-resolution structure and accompanying functional analyses have revealed the molecular mechanism of RNA-guided DNA targeting by Cas9, thus paving the way for the rational design of new, versatile genome-editing technologies. Japan. Science and Technology Agency (PRESTO) Japan Society for the Promotion of Science National Institutes of Health (U.S.) (NIH Director’s Pioneer Award (5DP1-MH100706)) |
| وصف الملف: | application/pdf |
| تدمد: | 0092-8674 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c372b4a9e4131272a04a18f8e6e6bdc3Test https://doi.org/10.1016/j.cell.2014.02.001Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....c372b4a9e4131272a04a18f8e6e6bdc3 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 00928674 |
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