Conformational control of DNA target cleavage by CRISPR-Cas9.


:Cas9 is an RNA-guided DNA endonuclease that targets foreign DNA for destruction as part of a bacterial adaptive immune system mediated by clustered regularly interspaced short palindromic repeats (CRISPR). Together with single-guide RNAs, Cas9 also functions as a powerful genome engineering tool in plants and animals, and efforts are underway to increase the efficiency and specificity of DNA targeting for potential therapeutic applications. Studies of off-target effects have shown that DNA binding is far more promiscuous than DNA cleavage, yet the molecular cues that govern strand scission have not been elucidated. Here we show that the conformational state of the HNH nuclease domain directly controls DNA cleavage activity. Using intramolecular Förster resonance energy transfer experiments to detect relative orientations of the Cas9 catalytic domains when associated with on- and off-target DNA, we find that DNA cleavage efficiencies scale with the extent to which the HNH domain samples an activated conformation. We furthermore uncover a surprising mode of allosteric communication that ensures concerted firing of both Cas9 nuclease domains. Our results highlight a proofreading mechanism beyond initial protospacer adjacent motif (PAM) recognition and RNA-DNA base-pairing that serves as a final specificity checkpoint before DNA double-strand break formation.






Sternberg SH,LaFrance B,Kaplan M,Doudna JA




Has Abstract


2015-11-05 00:00:00
















  • Rate of turnover of structural variants in the rDNA gene family of Drosophila melanogaster.

    abstract::A high degree of polymorphism for the length and copy number of rDNA spacers, in both the X and Y chromosome clusters, has been found in a wild population of Drosophila melanogaster. The genetic behaviour of rDNA structural variants in separate and mixed populations derived from isofemale lines suggest that they are n...


    pub_type: 杂志文章


    authors: Coen ES,Thoday JM,Dover G

    更新日期:1982-02-18 00:00:00

  • DNA double-strand breaks activate a multi-functional genetic program in developing lymphocytes.

    abstract::DNA double-strand breaks are generated by genotoxic agents and by cellular endonucleases as intermediates of several important physiological processes. The cellular response to genotoxic DNA breaks includes the activation of transcriptional programs known primarily to regulate cell-cycle checkpoints and cell survival....


    pub_type: 杂志文章


    authors: Bredemeyer AL,Helmink BA,Innes CL,Calderon B,McGinnis LM,Mahowald GK,Gapud EJ,Walker LM,Collins JB,Weaver BK,Mandik-Nayak L,Schreiber RD,Allen PM,May MJ,Paules RS,Bassing CH,Sleckman BP

    更新日期:2008-12-11 00:00:00

  • Induction of gadd45beta by NF-kappaB downregulates pro-apoptotic JNK signalling.

    abstract::In addition to coordinating immune and inflammatory responses, NF-kappaB/Rel transcription factors control cell survival. Normally, NF-kappaB dimers are sequestered in the cytoplasm by binding to inhibitory IkappaB proteins, and can be activated rapidly by signals that induce the sequential phosphorylation and proteol...


    pub_type: 杂志文章


    authors: De Smaele E,Zazzeroni F,Papa S,Nguyen DU,Jin R,Jones J,Cong R,Franzoso G

    更新日期:2001-11-15 00:00:00

  • Violation of the incompressibility of liquid by simple shear flow.

    abstract::In standard fluid dynamics, the density change associated with flow is often assumed to be negligible, implying that the fluid is incompressible. For example, this has been established for simple shear flows, where no pressure change is associated with flow: there is no volume deformation due to viscous stress and ine...


    pub_type: 杂志文章


    authors: Furukawa A,Tanaka H

    更新日期:2006-09-28 00:00:00

  • Author Correction: Anti-tumour immunity controlled through mRNA m6A methylation and YTHDF1 in dendritic cells.

    abstract::In this Letter, a citation to 'Fig. 1e' has been corrected to 'Fig. 1d' in the sentence starting "By contrast, the anti-tumour response…". This has been corrected online. ...


    pub_type: 已发布勘误


    authors: Han D,Liu J,Chen C,Dong L,Liu Y,Chang R,Huang X,Liu Y,Wang J,Dougherty U,Bissonnette MB,Shen B,Weichselbaum RR,Xu MM,He C

    更新日期:2019-04-01 00:00:00

  • CD95's deadly mission in the immune system.

    abstract::Apoptosis in the immune system is a fundamental process regulating lymphocyte maturation, receptor repertoire selection and homeostasis. Thus, death by apoptosis is as essential for the function of lymphocytes as growth and differentiation. This article focuses on death receptor-associated apoptosis and the role of CD...


    pub_type: 杂志文章,评审


    authors: Krammer PH

    更新日期:2000-10-12 00:00:00

  • Topoisomerase II mediates meiotic crossover interference.

    abstract::Spatial patterning is a ubiquitous feature of biological systems. Meiotic crossovers provide an interesting example, defined by the classic phenomenon of crossover interference. Here we identify a molecular pathway for interference by analysing crossover patterns in budding yeast. Topoisomerase II plays a central role...


    pub_type: 杂志文章


    authors: Zhang L,Wang S,Yin S,Hong S,Kim KP,Kleckner N

    更新日期:2014-07-31 00:00:00

  • Valuation of consumption and sale of forest goods from a Central American rain forest

    abstract::Researchers recognize that society needs accurate and comprehensive estimates of the economic value of rain forests to assess conservation and management options. Valuation of forests can help us to decide whether to implement policies that reconcile the value different groups attach to forests. Here we have measured ...


    pub_type: 杂志文章


    authors: Godoy R,Wilkie D,Overman H,Cubas A,Cubas G,Demmer J,McSweeney K,Brokaw N

    更新日期:2000-07-06 00:00:00

  • Mechanism of transfer RNA maturation by CCA-adding enzyme without using an oligonucleotide template.

    abstract::Transfer RNA nucleotidyltransferases (CCA-adding enzymes) are responsible for the maturation or repair of the functional 3' end of tRNAs by means of the addition of the essential nucleotides CCA. However, it is unclear how tRNA nucleotidyltransferases polymerize CCA onto the 3' terminus of immature tRNAs without using...


    pub_type: 杂志文章


    authors: Xiong Y,Steitz TA

    更新日期:2004-08-05 00:00:00

  • Enkephalin-, VIP- and substance P-like immunoreactivity in the carotid body.

    abstract::The carotid body type I cell contains amines and has features, both morphological and cytochemical, which indicate that it may also produce a peptide. Many regulatory peptides are now known to be present in both central and peripheral tissues. In the periphery these neuropeptides occur in both classical endocrine (APU...


    pub_type: 杂志文章


    authors: Wharton J,Polak JM,Pearse AG,McGregor GP,Bryant MG,Bloom SR,Emson PC,Bisgard GE,Will JA

    更新日期:1980-03-20 00:00:00

  • Robotics: self-reproducing machines.

    abstract::Self-reproduction is central to biological life for long-term sustainability and evolutionary adaptation. Although these traits would also be desirable in many engineered systems, the principles of self-reproduction have not been exploited in machine design. Here we create simple machines that act as autonomous modula...


    pub_type: 杂志文章


    authors: Zykov V,Mytilinaios E,Adams B,Lipson H

    更新日期:2005-05-12 00:00:00

  • CD95 promotes tumour growth.

    abstract::CD95 (also called Fas and APO-1) is a prototypical death receptor that regulates tissue homeostasis mainly in the immune system through the induction of apoptosis. During cancer progression CD95 is frequently downregulated or cells are rendered apoptosis resistant, raising the possibility that loss of CD95 is part of ...


    pub_type: 杂志文章


    authors: Chen L,Park SM,Tumanov AV,Hau A,Sawada K,Feig C,Turner JR,Fu YX,Romero IL,Lengyel E,Peter ME

    更新日期:2010-05-27 00:00:00

  • Developmental regulation of NMDA receptor-mediated synaptic currents at a central synapse.

    abstract::The central nervous system has extraordinary plasticity in early life. This is thought to involve N-methyl-D-aspartate (NMDA) receptors which, along with the non-NMDA receptors, mediate fast excitatory synaptic transmission. Although NMDA receptors may be transiently enhanced early in life, it has not been possible to...


    pub_type: 杂志文章


    authors: Hestrin S

    更新日期:1992-06-25 00:00:00

  • Influence of the Gulf Stream on the troposphere.

    abstract::The Gulf Stream transports large amounts of heat from the tropics to middle and high latitudes, and thereby affects weather phenomena such as cyclogenesis and low cloud formation. But its climatic influence, on monthly and longer timescales, remains poorly understood. In particular, it is unclear how the warm current ...


    pub_type: 杂志文章


    authors: Minobe S,Kuwano-Yoshida A,Komori N,Xie SP,Small RJ

    更新日期:2008-03-13 00:00:00

  • Synthetic lethal metabolic targeting of cellular senescence in cancer therapy.

    abstract::Activated oncogenes and anticancer chemotherapy induce cellular senescence, a terminal growth arrest of viable cells characterized by S-phase entry-blocking histone 3 lysine 9 trimethylation (H3K9me3). Although therapy-induced senescence (TIS) improves long-term outcomes, potentially harmful properties of senescent tu...


    pub_type: 杂志文章


    authors: Dörr JR,Yu Y,Milanovic M,Beuster G,Zasada C,Däbritz JH,Lisec J,Lenze D,Gerhardt A,Schleicher K,Kratzat S,Purfürst B,Walenta S,Mueller-Klieser W,Gräler M,Hummel M,Keller U,Buck AK,Dörken B,Willmitzer L,Reimann M,

    更新日期:2013-09-19 00:00:00

  • Visible-frequency hyperbolic metasurface.

    abstract::Metamaterials are artificial optical media composed of sub-wavelength metallic and dielectric building blocks that feature optical phenomena not present in naturally occurring materials. Although they can serve as the basis for unique optical devices that mould the flow of light in unconventional ways, three-dimension...


    pub_type: 杂志文章


    authors: High AA,Devlin RC,Dibos A,Polking M,Wild DS,Perczel J,de Leon NP,Lukin MD,Park H

    更新日期:2015-06-11 00:00:00

  • Polymer-supported membranes as models of the cell surface.

    abstract::Lipid-bilayer membranes supported on solid substrates are widely used as cell-surface models that connect biological and artificial materials. They can be placed either directly on solids or on ultrathin polymer supports that mimic the generic role of the extracellular matrix. The tools of modern genetic engineering a...


    pub_type: 杂志文章,评审


    authors: Tanaka M,Sackmann E

    更新日期:2005-09-29 00:00:00

  • GTP binding and hydrolysis by the signal recognition particle during initiation of protein translocation.

    abstract::The signal recognition particle (SRP) consists of one RNA and six protein subunits. The N-terminal domain of the 54K subunit contains a putative GTP-binding site, whereas the C-terminal domain binds signal sequences and SRP RNA. Binding of SRP to the signal sequence as it emerges from the ribosome creates a cytosolic ...


    pub_type: 杂志文章


    authors: Miller JD,Wilhelm H,Gierasch L,Gilmore R,Walter P

    更新日期:1993-11-25 00:00:00

  • Reduction of disulphide bonds unmasks potent antimicrobial activity of human β-defensin 1.

    abstract::Human epithelia are permanently challenged by bacteria and fungi, including commensal and pathogenic microbiota. In the gut, the fraction of strict anaerobes increases from proximal to distal, reaching 99% of bacterial species in the colon. At colonic mucosa, oxygen partial pressure is below 25% of airborne oxygen con...


    pub_type: 杂志文章


    authors: Schroeder BO,Wu Z,Nuding S,Groscurth S,Marcinowski M,Beisner J,Buchner J,Schaller M,Stange EF,Wehkamp J

    更新日期:2011-01-20 00:00:00

  • The Ndc80 kinetochore complex forms oligomeric arrays along microtubules.

    abstract::The Ndc80 complex is a key site of regulated kinetochore-microtubule attachment (a process required for cell division), but the molecular mechanism underlying its function remains unknown. Here we present a subnanometre-resolution cryo-electron microscopy reconstruction of the human Ndc80 complex bound to microtubules...


    pub_type: 杂志文章


    authors: Alushin GM,Ramey VH,Pasqualato S,Ball DA,Grigorieff N,Musacchio A,Nogales E

    更新日期:2010-10-14 00:00:00

  • Ephrin-B2 regulates VEGFR2 function in developmental and tumour angiogenesis.

    abstract::The formation and guidance of specialized endothelial tip cells is essential for both developmental and pathological angiogenesis. Notch-1 signalling regulates the generation of tip cells, which respond to gradients of vascular endothelial growth factor (VEGF-A). The molecular cues and signalling pathways that control...


    pub_type: 杂志文章


    authors: Sawamiphak S,Seidel S,Essmann CL,Wilkinson GA,Pitulescu ME,Acker T,Acker-Palmer A

    更新日期:2010-05-27 00:00:00

  • Elevated UV-B radiation reduces genome stability in plants.

    abstract::Long-term depletion of the stratospheric ozone layer contributes to an increase in terrestrial solar ultraviolet-B radiation. This has deleterious effects on living organisms, such as DNA damage. When exposed to elevated ultraviolet-B radiation (UV-B; 280-315 nm), plants display a wide variety of physiological and mor...


    pub_type: 杂志文章


    authors: Ries G,Heller W,Puchta H,Sandermann H,Seidlitz HK,Hohn B

    更新日期:2000-07-06 00:00:00

  • Dephosphorylation and activation of a p34cdc2/cyclin B complex in vitro by human CDC25 protein.

    abstract::Oocytes arrested in the G2 phase of the cell cycle contain a p34cdc2/cyclin B complex which is kept in an inactive form by phosphorylation of its p34cdc2 subunit on tyrosine, threonine and perhaps serine residues. The phosphatase(s) involved in p34cdc2 dephosphorylation is unknown, but the product of the fission yeast...


    pub_type: 杂志文章


    authors: Strausfeld U,Labbé JC,Fesquet D,Cavadore JC,Picard A,Sadhu K,Russell P,Dorée M

    更新日期:1991-05-16 00:00:00

  • Random quasi-phase-matching in bulk polycrystalline isotropic nonlinear materials.

    abstract::Three-wave mixing in nonlinear materials--the interaction of two light waves to produce a third--is a convenient way of generating new optical frequencies from common laser sources. However, the resulting optical conversion yield is generally poor, because the relative phases of the three interacting waves change cont...


    pub_type: 杂志文章


    authors: Baudrier-Raybaut M,Haïdar R,Kupecek P,Lemasson P,Rosencher E

    更新日期:2004-11-18 00:00:00

  • Pharmacological activation of REV-ERBs is lethal in cancer and oncogene-induced senescence.

    abstract::The circadian clock imposes daily rhythms in cell proliferation, metabolism, inflammation and DNA damage response. Perturbations of these processes are hallmarks of cancer and chronic circadian rhythm disruption predisposes individuals to tumour development. This raises the hypothesis that pharmacological modulation o...


    pub_type: 杂志文章


    authors: Sulli G,Rommel A,Wang X,Kolar MJ,Puca F,Saghatelian A,Plikus MV,Verma IM,Panda S

    更新日期:2018-01-18 00:00:00

  • Progenitor identification and SARS-CoV-2 infection in human distal lung organoids.

    abstract::The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate the investigation of pathologies such as interstitial lung disease, cancer and coronavirus disease 2019 (COVID-19) pneumonia caused by severe ac...


    pub_type: 杂志文章


    authors: Salahudeen AA,Choi SS,Rustagi A,Zhu J,van Unen V,de la O SM,Flynn RA,Margalef-Català M,Santos AJM,Ju J,Batish A,Usui T,Zheng GXY,Edwards CE,Wagar LE,Luca V,Anchang B,Nagendran M,Nguyen K,Hart DJ,Terry JM,Belgrad

    更新日期:2020-12-01 00:00:00

  • A microRNA controlling left/right neuronal asymmetry in Caenorhabditis elegans.

    abstract::How left/right functional asymmetry is layered on top of an anatomically symmetrical nervous system is poorly understood. In the nematode Caenorhabditis elegans, two morphologically bilateral taste receptor neurons, ASE left (ASEL) and ASE right (ASER), display a left/right asymmetrical expression pattern of putative ...


    pub_type: 杂志文章


    authors: Johnston RJ,Hobert O

    更新日期:2003-12-18 00:00:00

  • Spermiogenesis deficiency and germ-cell apoptosis in CREM-mutant mice.

    abstract::Spermiogenesis is a complex process by which postmeiotic male germ cells differentiate into mature spermatozoa. This process involves remarkable structural and biochemical changes including nuclear DNA compaction and acrosome formation. Transcription activator CREM (cyclic AMP-responsive element modulator) is highly e...


    pub_type: 杂志文章


    authors: Nantel F,Monaco L,Foulkes NS,Masquilier D,LeMeur M,Henriksén K,Dierich A,Parvinen M,Sassone-Corsi P

    更新日期:1996-03-14 00:00:00

  • Relatives of rubella virus in diverse mammals.

    abstract::Since 1814, when rubella was first described, the origins of the disease and its causative agent, rubella virus (Matonaviridae: Rubivirus), have remained unclear1. Here we describe ruhugu virus and rustrela virus in Africa and Europe, respectively, which are, to our knowledge, the first known relatives of rubella viru...


    pub_type: 杂志文章


    authors: Bennett AJ,Paskey AC,Ebinger A,Pfaff F,Priemer G,Höper D,Breithaupt A,Heuser E,Ulrich RG,Kuhn JH,Bishop-Lilly KA,Beer M,Goldberg TL

    更新日期:2020-10-01 00:00:00

  • Calculation of the axion mass based on high-temperature lattice quantum chromodynamics.

    abstract::Unlike the electroweak sector of the standard model of particle physics, quantum chromodynamics (QCD) is surprisingly symmetric under time reversal. As there is no obvious reason for QCD being so symmetric, this phenomenon poses a theoretical problem, often referred to as the strong CP problem. The most attractive sol...


    pub_type: 杂志文章


    authors: Borsanyi S,Fodor Z,Guenther J,Kampert KH,Katz SD,Kawanai T,Kovacs TG,Mages SW,Pasztor A,Pittler F,Redondo J,Ringwald A,Szabo KK

    更新日期:2016-11-03 00:00:00