groups期刊最新论文

近五年作为通讯作者，共发表了研究论文200余篇，其中被SCI收录180余篇，主要发表在国际著名的Adv. Funct. Mater.、Chem: .、Chem. Commun.、J. Mater. Chem.、 J. Phys. Chem. B、J. Phys. Chem. C、J. Comput. Chem.、Inorg. Chem.、Appl. Catal. B、Nanotechnology、J. Phys.: Condensed Matter等学术期刊上，影响因子在以上的论文有50余篇，其中影响因子在以上8篇。成果被SCI收录期刊正面引用1400余次。出版专著两部，美国科学出版社邀请为纳米科学技术大百科全书撰写了完整的一章。申请发明专利14项。已发表和已录用的主要论文清单如下（以下均为SCI期刊收录论文，*代表通讯联系人）·Pan Qing-Jiang,Guo Yuan-Ru,Li Li,Fu Hong-Gang*,and Zhang Hong-Xing “Structures,Spectroscopic Properties and Redox Potentials of Quaterpyridyl Ru（Ⅱ） Photosensitizer and its Derivatives for Solar Energy Cell: A Density Functional Study” Physical Chemistry Chemical Physics 2011,DOI:.·Gao Xiao-Qin,Pan Qing-Jiang,Li Li,Guo Yuan-Ru,Zhang Hong-Xing,Fu Hong-Gang* “Structures and Spectroscopic Properties of Ruthenium Phenanthroline Solar-Cell Sensitizers: A Computational Study” Chemical Physics Letters,2011,506,146-151.·Zhou W,Sun FF,Pan K,Tian GH,Fu HG*. Well-ordered large-pore mesoporous anatase TiO2 with remarkably high thermal stability and improved crystallinity: Preparation,characterization and photocatalytic performance. Advanced Functional Materials,2011,21,1922-1930..·Jiang Baojiang,Tian CG,Fu Honggang*,et al. In-Situ Growth of TiO2 in Interlayers of Expanded Graphite for the Fabrication of TiO2-Graphene with Enhanced Photocatalytic Activity. Chemistry-A European Journal,2011,Accepted,.· Li Sun,Chungui Tian,Lei Wang,Jinlong Zou,Guang Mu,Honggang Fu*,Magnetically Separable Porous Graphitic Carbon with Large Surface Area as Excellent Adsorbents for Metal Ions and Dye. Journal of Materials Chemistry,2011,2011,21,7232-7239.· Zhao H,Yang J,Wang L,Tian CG,Jiang BJ,Fu HG*. Fabrication of palladium nanoparticles/graphene nanosheets hybrid via sacrifice of copper template and its application in catalytic oxidation of formic acid. Chemical Communications,2011,47,2014-2016.· Yang J,Tian CG,Wang L,Fu HG*. An effective strategy to small-sized and high-dispersed palladium nano particles supported on graphene with excellent performance for formic acid oxidation. Journal of Materials Chemistry,2011,21,3384-3390.· Tian GH,Chen YJ,Zhou W,Ren ZY,Tian CG,Fu HG*. Facile solvothermal synthesis of hierarchical flower-like Bi2MoO6 hollow spheres as high performance visible-light driven photocatalysts. Journal of Materials Chemistry,2011,21,887 – 892.· Hu ZF,Shen PK*,Fu HG*,et al. Oxygen Reduction Electrocatalysis Enhanced by Nanosized Cubic Vanadium Carbide. Electrochemistry Communications,2011,Accepted,ELECOM 3881.· Tian Guohui,Chen Yajie,Zhou Wei,Pan Kai,Huang Xu-ri and Fu Honggang *,3D hierarchical flower-like TiO2 nanostructure: morphology control and its photocatalytic property. CrstEngComm,2011,13,2994.· Dong Youzhen,Pan Kai,Zhou Wei,Pan Qingjiang,Xie Tengfeng,Wang Dejun,Fu Honggang*,Dye-sensitized solar cells based on TiO2-B nanobelt/TiO2 nanoparticle sandwich-type photoelectrodes with controllable nanobelt length. Dalton Transactions,2011,40,3808–3814.· Cai ZC,Tian CG,Wang L,Zhou W,Wang BL,Fu HG*. One-pot synthesis of silver particle aggregation as highly active SERS substrate. Journal of Raman Spectroscopy,2011,1,5-11.· Tian CG,Li W,Fu HG*,et al. Glucose-mediated solution-solid route for easy synthesis of Ag/ZnO particles with superior photocatalytic activity and photostability，Journal of Alloys and Compounds，2011,509,6935-6941.· Tian CG,Li W,Fu HG*,et al. Controllable Fabrication of Various ZnO Micro/nanostructures from a Wire-like Zn-EG-AC Precursor via a Facile Solution-based Route，Materials Research Bulletin，2011，MRB5116.· Zhao H,Fu HG,Tian CG,et shape-controlled synthesis of palladium nanostructures on copper for promising Surface-enhanced Raman scattering,MATERIALS LETTERS,2010,64,2255-2257· Zhao H,Fu HG,Tian CG,et al. Fabrication of silver nanoparticles/single-walled carbon nanotubes composite for surface-enhanced Raman scattering,JOURNAL OF COLLOID AND INTERFACE SCIENCE,2010,351,343-347.· Jiang BJ,Tian CG,Fu HG*,et al. Facile Fabrication of High Quality Graphene from Expandable Graphite: Simultaneous Exfoliation and Reduction. Chemical Communication. 2010,46,4920 - 4922.· Feng SS,Ren ZY,Fu HG*,et al. Synthesis and application of hollow magnetic graphitic carbon microspheres with/without TiO2 nanoparticle layer on the surface. Chemical Communication. 2010,46,6276-6278.· Liu Y,Ren ZY,Fu HG*,et al. Synthesis and applications of graphite carbon sphere with uniformly distributed magnetic Fe3O4 nanoparticles (MGCSs) and MGCS@Ag,MGCS@TiO2. Journal of Material Chemistry 2010,20，4802- 4808.· Wang L,Tian CG,Fu HG*,et al. Mass production of graphene via an in situ self-generating template route and its promoted activity as electrocatalytic support for methanol electroxidization. Journal of Physical Chemistry C 2010,114 （19），8727–8733.· Qu Y,Zhou W,Fu HG*,et al. Hierarchical anatase TiO2 porous nanopillars with high crystallinity and controlled length: An effective candidate for dye-sensitized solar-cells. Physical Chemistry Chemical Physics 2010,12,9205 - 9212.· Wang BL,Tian CG,Fu HG,et al. Chitosan: a green carbon source for the synthesis of graphitic nanocarbon,tungsten carbide and graphitic nanocarbon/tungsten carbide composites. Nanotechnology 21 （2010） 025606 （9pp).· Li YJ,LV RJ,Fu HG*,et al. Fabrication and evaluation of chiral monolithic column modified by β-cyclodextrin derivatives. Talanta 80 （2010） 1378–1382· Zhou W,Pan K,Fu HG*,et al. Photodegradation of organic contamination in wastewaters by bondingTiO2/single-walled carbon nanotube composites with enhanced photocatalytic activity. Chemosphere 81 （2010） 555–561.· Guohui Tian,Chen Yajie,Pan Kai,Fu Honggang*，Efficient visible light-induced degradation of phenol on N-doped anatase TiO2 with large surface area and high crystallinity. Applied Surface Science 2010,256,3740-3745.· Tian CG,Li W,Fu HG*,et al. One-pot Synthesis of the Ag Nanoparticles modified ZnO Microspheres in Ethylene Glycol Medium and Their Enhanced Photocatalytic Performance. Journal of Solid State Chemistry,2010,183,2720-2725.· Zhou W,Pan K,Fu HG*,et al. The sandwich structure electrodes based on wire-likeTiO2–β-cyclodextrin–SWCNT composite for dye-sensitized solar cells. Journal of Photochemistry and Photobiology A: Chemistry 2009,207,306–310.· Tian XQ,Wan LJ,Fu HG*,et al. Facile synthesis of mesoporous ZnAl2O4 thin films through the evaporation-induced self-assembly method. Journal of Alloys and Compounds 2009,488,320–324.· Wang RH,Tian CG,Fu HG*,et al. In situ simultaneous synthesis of WC/graphitic carbon nanocomposite as a highly efficient catalyst support for DMFCw. Chemical Communication 2009,3104-3106.· Pan QJ,Fu HG*,Zhang HX*,et al. Theoretical Studies on Metal−Metal Interaction,Excited States,and Spectroscopic Properties of Binuclear Au−Au,Au−Rh,and Rh−Rh Complexes with Diphosphine Ligands: Buildup of Complexity from Monomers to Dimers. Inorganic Chemistry,2009,48,2844–2854.· Zhou W,Pan K,Fu HG*,et al. Solar-induced Self-assembly of TiO2-β-cyclodextrin-MWCNT Composite Wires. Physical Chemistry Chemical Physics 2009,11,1713-1718.· Kang CH,Jing LQ*,Fu HG*,et al. Mesoporous SiO2-Modified Nanocrystalline TiO2 with High Anatase Thermal Stability and Large Surface Area as Efficient Photocatalyst. Journal of Physical Chemistry C 2009,113,1006–1013.· Wang BL,Tian CG,Fu HG*,et al. A simple and large-scale strategy for the preparation of Ag nanoparticles supported on resin-derived carbon and their antibacterial properties. Nanotechnology 2009,20,025603（7pp）· Tian GH,Fu HG*,Jing LQ,et al. Synthesis and photocatalytic activity of stable nanocrystalline TiO2 with high crystallinity and large surface area. Journal of Hazardous Materials 2009,161,1122-1130.· Zhang GX,Yu HT*,Fu HG,et al. First-principles calculations of the stability and electronic properties of the PbTiO3 （110） polar surface. Journal of Computational Chemistry 2009,30,1785-1798.· Tian GH,Pan K,Fu HG*,et al. Enhanced photocatalytic activity of S-doped TiO2-ZrO2 nanoparticles under visible-light irradiation. Journal of Hazardous Materials 2009,166,939-944.· Pan K,Dong YZ,Fu HG*,et al. TiO2-B narrow nanobelt/TiO2 nanoparticle composite photoelectrode for dye-sensitized solar cells. Electrochimica Acta 2009,54,7350-7356.· Kan K,Fu HG,Shi KY*,et al. Amidation of single-walled carbon nanotubes by a hydrothermal process for the electrooxidation of nitric oxide. Nanotechnology 2009,20,185502 （7pp).· Wang L,Tian CG,Fu HG*,et al. Controllable Synthesis of Graphitic Carbon Nanostructures from Ion-Exchange Resin-Iron Complex via Solid-State Pyrolysis Process. Chemical Communication 2008,5411-5413.· Zhou W,Fu HG*,Pan K,et al. Mesoporous TiO2/α-Fe2O3: Bifunctional Composites for Effective Elimination of Arsenite Contamination through Simultaneous Photocatalytic Oxidation and Adsorption. Journal of Physical Chemistry C 2008,112,19584-19589.· Liu KS,Fu HG*,Xie Y,et al. Assembly of β-Cyclodextrins Acting as Molecular Bricks onto the Multiwall Carbon of Physical Chemistry C 2008,112,951-957.· Tian GH,Fu HG*,Jing LQ,et al. Preparation and Characterization of Stable Biphase TiO2 Photocatalyst with High Crystallinity,Large Surface Area and Enhanced Photoactivity. Journal of Physical Chemistry C 2008,112,3083-3089.· Zhou W,Liu KS,Fu HG*,et al. Multi-modal mesoporous TiO2–ZrO2 composites with high photocatalytic activity and hydrophilicity. Nanotechnology 2008,19,035610（7pp).· Pan QJ,Zhou X,Fu HG*,et al，“Isovalent Gold(I),-（Ⅱ），and -（Ⅲ） and Mixed-Valent Gold(I)/Gold（Ⅲ） Phosphorus Ylide Complexes. Combined ab Initio and Density Functional Study of Electronic Structures and Spectroscopic Properties” Organometallics 2008,27,2474–2482.· Wan LJ,Fu HG*,Shi KY,et al. Facile synthesis of ordered nanocrystalline alumina thin films with tunable mesopore structures. Microporous and Mesoporous Materials 2008,115（3），301-307.· Pan QJ，Zhou X,Fu HG*,et al. A theoretical probe on the ground- and excited-state properties of heterobinuclear Au–Pt complex with phosphine ligands. Comparison with analogous homobinuclear Au–Au and Pt–Pt complexes. Chemical Physics Letters 2008,453,7-12.· Jing LQ*,Li SD,Fu HG*,et al. Investigation on the electron transfer between anatase and rutile in nano-sized TiO2 by means of surface photovoltage technique and its effects on the photocatalytic activity. Solar Energy Materials & Solar Cells 2008,92,1030– 1036.· Wan LJ,Fu HG*,Shi KY,et al. Facile synthesis of iron oxide with wormlike morphology and their application in water treatment. Journal of Solid State Chemistry 2008,181（4），735-740.· Chi YJ,Fu HG*,Qi LH,et al. Preparation and photoelectric performance of ITO/TiO2/CdS composite thin films. Journal of Photochemistry and Photobiology A: Chemistry 2008，195（2-3），357-363.· Xie Y,Yu HT,Fu HG*,et al. Lattice dynamics investigation on different transition behaviors of cubic BaTiO3 and SrTiO3 by first-principles calculatoins. J. Phys.: Condens. Matter,2008,20,215215 （8pp)· Wan LJ,Fu HG*,Shi KY,et al. Simple synthesis of mesoporous alumina thin Letters 2008,62（10-11），1525-1527.· Song S,Jing LQ*,Fu HG*,et al. Superhydrophilic anatase TiO2 film with the micro- and nanometer-scale hierarchical surface structure. Materials Letters 2008,62,3503–3505.· Zhu YJ,Yuan FL*,Fu HG,et al. Direct NO decomposition over La2-xBaxNiO4 catalysts containing BaCO3 phase. Appl. Catal B-Environ 2008,82,255-263.· Li MX,Zhang HX*,Fu HG,et al. Theoretical Studies of Electronic Structuresand Spectroscopic Properties of Ru Complexes. Inorg. Chem 2008 47（7），2312-2324.· Xie Y,Yu HT,Fu HG*,A First-Principles Investigation of Stability and Structural Properties of the BaTiO3 （110） Polar Surface. Journal of Physical Chemistry C,2007,111,6343-6349.· Kan W,Yu HT,Fu HG*,et al. Theoretical investigation on the protonation reactions and products of the stable [N,C,C,S] isomers,Journal of Computational Chemistry,2007,28,2472-2482.· Zhao YL,Yu HT*,Fu HG,et al. Combined DFT,QCISD(T),and G2 mechanism investigation for the reactions of carbon monophosphide CP with unsaturated hydrocarbons allene CH2CCH2 and methylacetylene CH3CCH,Journal of Computational Chemistry,2007,28,1221-1233.· Pan QJ,Zhang HX*,Fu HG,et al. Electronic Structures and Spectroscopic Properties of Mono- and Binuclear d8 Complexes: a Theoretical Exploration on Promising Phosphorescent Materials. Journal of Physical Chemistry A,2007,111（2）; 287-294.· Pan QJ,Zhou X,Fu HG*,et al. Theoretical studies on spectroscopic properties of binuclear palladium（Ⅱ） halide with phosphine ligands Journal of Photochemistry and Photobiology A: Chemistry,2007,188（2-3），287-292.· Xie Y,Fu HG*,Yu HT,et al. A first-principles investigation into the ferroelectric and antiferrodistortive instabilities of cubic SrTiO3,Journal of Physics: Condensed Matter,2007,19,506213（9pp)· Pan QJ,Zhou X,Fu HG*,et alStructures and electronic spectra of [Pt2(P2O5H2）4X2]4– (X = Cl,Br and I): a comparative study of ab initio and density functional theory Inorganic Chemistry Communications 2007,10,183–186.· Li SD,Jing LQ,Fu HG*,et al. Photoinduced charge property of nanosized perovskite-type LaFeO3 and its relationships with photocatalytic activity under visible irradiation. Materials Research Bulletin 2007,42,203-212.· Yang PP,Fu HG,Lin J*,et al. MCM-41 functionalized with YVO4: Eu3+: a novel drug delivery system. Nanotechnology 2007,18,235703（6pp).· Yang PP,Fu HG,Lin J*,et al. Luminescence functionalization of SBA-15 by YVO4: Eu3+ as a novel drug delivery system. Inorg. Chem. 2007,46,3202-3211.· Jing LQ,Fu HG*,Wang BQ,et al. Effects of Sn dopant on the photoinduced charge property and photocatalytic activity of TiO2 nanoparticles. Applied Catalysis B,2006,62,282-291.· Jing LQ,Xin BF,Fu HG*,et al. Effects of Surface Oxygen Vacancies on Photophysical and Photochemical Processes of Zn-Doped TiO2 Nanoparticles and Their Relationships. Journal of Physical Chemistry B2006,110,17860-17865.· Pan QJ,Fu HG*,Yu HT,et al. A Theoretical Insight Into Electronic Structures and Spectroscopic Properties of [Pt2(pop)4]4–，[Pt2(pcp)4]4– and Related Derivatives (pop = P2O5H22– and pcp = P2O4CH42–） Inorganic Chemistry，2006,45（21），8729–8735.· Liu KS,Zhou W,Fu HG*,et al. Influence of calcination temperatures on the photocatalytic activity and photo-induced hydrophilicity of wormhole-like mesoporous TiO2. Nanotechnology 2006,17,1363-1369.· Liu KS,Fu HG*,Shi KY,et al. Hydrophilicity and formation mechanism of large-pore mesoporous TiO2 thin films with tunable pore diameters. Nanotechnology 2006,17,3641-3648.· Pan QJ,Zhang HX*,Fu HG,et al “Theoretical Studies on Metal–Metal Interaction and Intrinsic 1,3[σ*(d）σ（s/p)] Excited States of Binuclear d10 Complexes with Bridging Phosphine Ligands” European Journal of Inorganic Chemistry 2006 （5），1050-1059.· Jing LQ,Wang DJ,Fu HG*,et al. Effects of noble metal modification on surface oxygen composition,charge separation and photocatalytic activity of ZnO nanoparticles. Journal of Molecular Catalysis A 2006,244,193-200.· Pan QJ,Fu HG*,Yu HT,et al Substituent effect on the structure and luminescence of binuclear Au(I) complexes: An ab initio study. Chemical Physics Letters 2006,426（4-6），257-262.· Jing LQ,Qu YC,Fu HG*,et al. Review of photoluminescence performance of nano-sized semiconductor materials and its relationships with photocatalytic activity. Solar Energy Materials & Solar Cell 2006,90,1773-1787.· Pan QJ,Fu HG*,Yu HT,et al Spectroscopic Properties of Mono- and Binuclear Platinum（Ⅱ） Alkynyl Complexes with Phosphine Ligands: A Theoretical StudyInorganica Chimica Acta 2006,359（10），3306–3314.· Wang BQ,Jing LQ,Fu HG*,et al. Enhancement of the photocatalytic activity of TiO2 nanoparticles by surface-capping DBS groups. Applied Surface Science 2006,252,2817-2825.· Liu KS,Zhang ML,Fu HG*,et al. Uniform TiO2 thin films with anatase nanocrystallites synthesized through evaporation-induced assembly. Journal of Non-Crystalline Solids 2006,352,2284–2287.· Liu KS,Fu HG*,Shi KY,et al. Preparation of large-pore mesoporous nanocrystalline TiO2 thin films with tailored pore diameters. Journal of Physical Chemistry B 2005,109 （40），18719-18722.· Shi KY,Chi YJ,Fu HG*,et al. Controlled growth of mesostructured crystalline iron oxide nanowires and Fe-filled carbon nanotube arrays templated by mesoporous silica SBA-16 film. Journal of Physical Chemistry B2005,109 （7），2546-2551.· Xin BF,Jing LQ,Fu HG*,et al. Effects of simultaneously doped and deposited Ag on the photocatalytic activity and surface states of TiO2. Journal of Physical Chemistry B2005,109,2805-2809.· Liu KS,Zhang ML,Fu HG*,et al. Preparation,characterization,and photo-induced hydrophilicity of nanocrystalline anatase thin films synthesized through evaporation-induced assembly. Nanotechnology 2005,16,3006-3011.· Xin BF,Ren ZY,Fu HG*,et al. Photocatalytic activity and interfacial carrier transfer of Ag–TiO2 nanoparticle films. Applied Surface Science 2005,252,2050–2055.· Liu KS,Zhang ML,Fu HG*,et al. Large pore mesoporous nanocrystalline titania thin films synthesized through evaporation-induced self-assembly. Materials Letters 2005,59,3308 -3310.

从简单地剪切致病基因，到开发出不再传播疾病的工程动物，基因编辑技术已经释放出巨大的潜力。随着研究的深入，科学界还发现，除了编辑具有遗传讯息的DNA片段，编辑RNA可以在不改变基因组的情况下，帮助调整基因表达方式，此外，RNA的寿命是相对短暂的，这也意味着它的变化是可以逆转的，从而避免基因工程中的巨大风险。

2017年10月，来自Broad研究所的张锋研究团队在《自然》期刊上发表了题为“RNA targeting with CRISPR-Cas13”的文章，首次将CRISPR-Cas13系统公之于众，证实了CRISPR-Cas13可以靶向哺乳动物细胞中的RNA。仅仅时隔三周，又一篇名为“RNA editing with CRISPR-Cas13”的力作发表于《科学》期刊。在该研究中，张锋研究团队再次展示了这一RNA编辑系统，能有效地对RNA中的腺嘌呤进行编辑。

在CRISPR出现之前，RNAi是调节基因表达的理想方法。但是Cas13a酶一大优势在于更强的特异性，而且这种本身来自细菌的系统对哺乳动物细胞来说，并不是内源性的，因此不太可能干扰细胞中天然的转录。相反，RNAi利用内源性机制进行基因敲除，对本身的影响较大。但CRISPR-Cas13系统还有一个重要的问题，Cas13a酶本质上是一种相对较大的蛋白质，因此很难被包装到靶组织中，这也可能成为RNA编辑技术临床应用的一大障碍。

2018年3月16日，一项发表在《细胞》期刊的重磅成果为RNA编辑技术带来一大步飞跃，来自美国Salk研究所的科学家利用全新的CRISPR家族酶扩展了RNA编辑能力，并将这个新系统命名为“CasRx”。

CasRx（品红色）在人类细胞核中靶向RNA（灰色），Salk研究所

“生物工程师就像自然界的侦探一样，在DNA模式中寻找线索来帮助解决遗传疾病。CRISPR彻底改变了基因工程，我们希望将编辑工具从DNA扩展到RNA。”研究领导者Patrick Hsu博士表示，“RNA信息是许多生物过程的关键介质。在许多疾病中，这些RNA信息失去了平衡，因此直接靶向RNA的技术将成为DNA编辑的重要补充。”

除了高效性且无明显脱靶效应，新系统的一个关键特征是其依赖于一种比以前研究中物理尺寸更小的酶。 这对RNA编辑技术至关重要，这使得该编辑工具能够更容易被包装到病毒载体，并进入细胞进行RNA编辑。来自东京大学的科学家Hiroshi Nishimasu并未参与这项研究，他表示：“在这项研究中，研究人员发现了一种较Cas13d更加‘紧凑’的酶CasRx。从基础研究到治疗应用，我认为CasRx将成为非常有用的工具。”

此外，在这项研究中，研究人员还展示了利用这种新型RNA编辑系统来纠正RNA过程的能力。他们将CasRx包装到病毒载体中，并将其递送到利用额颞叶痴呆（FTD）患者干细胞中培养的神经细胞，最终使tau蛋白水平恢复到健康水平上，有效率达到80%。

Patrick Hsu博士最后说道：“基因编辑技术通过对DNA的切割带来基因序列的改变。在经过基因编辑的细胞中，其效果是永久的。虽然基因编辑技术能够很好地将基因完全关闭，但对调节基因的表达上并不那么优秀。展望未来，这一最新工具将在RNA生物学研究中发挥重要作用，并有望在未来凭借该技术对RNA相关疾病进行治疗。”

该研究探索了Cas13d家族蛋白CasRx敲低目的基因的最佳sgRNA组合，通过尾静脉注射质粒的方式，将CasRx系统和靶向Pten基因的sgRNA导入到小鼠肝脏细胞中，成功在小鼠肝脏中实现了Pten的高效沉默。

3月18日，《蛋白质与细胞》期刊在线发表了《Cas13d介导的肝脏基因表达下调对代谢功能的调控》的研究论文，该研究由中科院脑科学与智能技术卓越创新中心（神经科学研究所）、上海脑科学与类脑研究中心、神经科学国家重点实验室杨辉研究组和上海科技大学生命科学与技术学院黄鹏羽研究组合作完成。该研究探索了Cas13d家族蛋白CasRx敲低目的基因的最佳sgRNA组合，通过尾静脉注射质粒的方式，将CasRx系统和靶向Pten基因的sgRNA导入到小鼠肝脏细胞中，成功在小鼠肝脏中实现了Pten的高效沉默，证实了CasRx系统在成体动物体内也具有靶向沉默RNA的活性，通过增强下游蛋白AKT的磷酸化，影响了糖脂代谢相关基因的表达。同时，利用AAV递送CasRx和靶向Pscsk9的sgRNA到小鼠肝脏，有效降低了肝脏中PCSK9的蛋白表达，以及小鼠血液中的胆固醇水平。这为治疗后天性的代谢疾病提供了新方案。

同时，杨辉研究组与上海交通大学医学院附属上海第一人民医院孙晓东研究组合作，也探究了CasRx预防严重的眼部疾病——年龄相关性黄斑变性（AMD）的可能性，研究人员发现在体内使用CasRx敲低Vegfa的mRNA可以显著减少AMD小鼠模型中脉络膜新血管形成（CNV）的面积，验证了将RNA靶向的CRISPR系统用于治疗应用的潜力。相关研究论文《CasRx介导的RNA靶向策略可防止年龄相关的黄斑变性的小鼠模型中的脉络膜新生血管形成》3月3日在《国家科学评论》在线发表。

近年来，CRISPR/Cas9技术因其强大且便捷的DNA编辑能力而受到广泛关注。2016年，张锋实验室发现了一种新的Cas蛋白Cas13a，可以靶向RNA进行切割。之后人们又陆续发现了靶向RNA的Cas13b, Cas13c。由于Cas13家族蛋白靶向RNA的特点，理论上在一些特定疾病的检测和治疗上具有独特优势，因而成为近年来的研究热点。2018年，加州大学伯克利分校Patrick Hsu实验室发现了Cas13d家族。他们发现与RNA干扰技术相比，Cas13d介导的基因沉默具有更高的特异性（与数百个shRNA脱靶相比，Cas13d没有脱靶）和敲除效率（Cas13d达到96％，shRNA达到65％）。而与Cas9介导的基因敲除技术相比，Cas13d介导的基因沉默不会改变基因组DNA，因此这种基因沉默是可逆的，从而对一些后天性疾病（如因不良生活习惯导致的高血脂等后天代谢性疾病）的治疗更有优势。其中Cas13d家族的CasRx蛋白由于体积小，效率高，被认为是在未来应用中最具有优势的Cas13蛋白。

此前的工作都在细胞水平证明了CasRx的高效性和特异性，杨辉研究组的这两篇文章则更进一步在动物体内证明了CasRx的活性，为临床提供了可能性。为证明CasRx在动物体内的活性，研究人员分别针对目的基因进行了sgRNA的体外筛选，然后采用尾静脉注射敲低Pten的质粒、尾静脉注射敲低Pcsk9的AAV8病毒、眼部注射敲低Vegfa的AAV病毒。对注射后的小鼠进行相应分析，分别得到Pten基因下调及其下游蛋白AKT的磷酸化上调，Pcsk9下调造成血清胆固醇下调；Vegfa下调显著减少AMD小鼠模型中脉络膜新血管形成（CNV）的面积。

2020年3月18日，《蛋白质与细胞》期刊在线发表了《Cas13d介导的肝脏基因表达下调对代谢功能的调控》的研究论文，该研究由中科院脑科学与智能技术卓越创新中心（神经科学研究所）、上海脑科学与类脑研究中心、神经科学国家重点实验室杨辉研究组和上海科技大学生命科学与技术学院黄鹏羽研究组合作完成。该研究探索了Cas13d家族蛋白CasRx敲低目的基因的最佳sgRNA组合，通过尾静脉注射质粒的方式，将CasRx系统和靶向 Pten 基因的sgRNA导入到小鼠肝脏细胞中，成功在小鼠肝脏中实现了 Pten 的高效沉默， 证实了CasRx系统在成体动物体内也具有靶向沉默RNA的活性， 通过增强下游蛋白AKT的磷酸化，影响了糖脂代谢相关基因的表达。同时，利用AAV递送CasRx和靶向 Pscsk9 的sgRNA到小鼠肝脏， 有效降低了肝脏中PCSK9的蛋白表达，以及小鼠血液中的胆固醇水平 。这为治疗后天性的代谢疾病提供了新方案。

同时，杨辉研究组与上海交通大学医学院附属上海第一人民医院孙晓东研究组合作，也 探究了CasRx预防严重的眼部疾病——年龄相关性黄斑变性（AMD）的可能性，研究人员发现在体内使用CasRx敲低 Vegfa的mRNA可以显著减少AMD小鼠模型中脉络膜新血管形成（CNV）的面积**，验证了将RNA靶向的CRISPR系统用于治疗应用的潜力。相关研究论文《CasRx介导的RNA靶向策略可防止年龄相关的黄斑变性的小鼠模型中的脉络膜新生血管形成》3月3日在《国家科学评论》在线发表。

近年来，CRISPR/Cas9技术因其强大且便捷的DNA编辑能力而受到广泛关注。2016年，张锋实验室发现了一种新的Cas蛋白Cas13a，可以靶向RNA进行切割。之后人们又陆续发现了靶向RNA的Cas13b, Cas13c。由于Cas13家族蛋白靶向RNA的特点，理论上在一些特定疾病的检测和治疗上具有独特优势，因而成为近年来的研究热点。2018年，加州大学伯克利分校Patrick Hsu实验室发现了Cas13d家族。他们发现与RNA干扰技术相比，Cas13d介导的基因沉默具有更高的特异性（与数百个shRNA脱靶相比， Cas13d没有脱靶）和敲除效率（Cas13d达到96％ ，shRNA达到65％）。而与Cas9介导的基因敲除技术相比， Cas13d介导的基因沉默不会改变基因组DNA，因此这种基因沉默是可逆的 ，从而对一些后天性疾病（如因不良生活习惯导致的高血脂等后天代谢性疾病）的治疗更有优势。其中Cas13d家族的CasRx蛋白由于体积小，效率高，被认为是在未来应用中最具有优势的Cas13蛋白。

此前的工作都在细胞水平证明了CasRx的高效性和特异性，杨辉研究组的这两篇文章则更进一步在动物体内证明了CasRx的活性，为临床提供了可能性 。为证明CasRx在动物体内的活性，研究人员分别针对目的基因进行了sgRNA的体外筛选，然后采用尾静脉注射敲低 Pten 的质粒、尾静脉注射敲低 Pcsk9 的AAV8病毒、眼部注射敲低 Vegfa 的AAV病毒。对注射后的小鼠进行相应分析，分别得到 Pten 基因下调及其下游蛋白AKT的磷酸化上调， Pcsk9 下调造成血清胆固醇下调； Vegfa 下调显著减少AMD小鼠模型中脉络膜新血管形成（CNV）的面积。

图1 CasRx介导的 Pten 体内体外的下调（ Protein & Cell ）

A.质粒示意图；细胞中 Pten 的下调；检测PTEN及AKT的表达； 与shRNA脱靶比较；E.尾静脉注射质粒示意图；.免疫荧光，qPCR，western分别检测 Pten 及p-AKT的表达

图2 血清胆固醇的调节以及 Pcsk9 的可逆调控（ Protein & Cell ）

A.针对 Pcsk9 的AAV8病毒注射示意图；B.肝组织中 Pcsk9 的表达量；C.血清 PCSK9 的表达量；D.血清胆固醇水平；.血清ALT和AST的测定；G.可逆调节注射示意图； H. Pcsk9 的动态调控。

图3 AAV介导CasRx减少了AMD小鼠模型中CNV的面积（National Science Review）

A.小鼠和人序列比较以及sgRNA示意图；.在293T和N2a细胞中敲低 Vegfa ；蛋白的表达；病毒质粒示意图；F.实验流程图；的mRNA表达水平；.激光烧伤之前或之后7天的 Vegfa mRNA水平；诱导3天后的VEGFA蛋白水平；K.激光烧伤7天后，用PBS或AAV-CasRx- Vegfa 注射的代表性CNV图像；面积统计。

2020 年 4 月 8 日， Cell 期刊在线发表了题为 《Glia-to-Neuron Conversion by CRISPR-CasRx Alleviates Symptoms of Neurological Disease in Mice》 的研究论文，该研究由中国科学院脑科学与智能技术卓越创新中心（神经科学研究所）、上海脑科学与类脑研究中心、神经科学国家重点实验室 杨辉 研究组完成。

该项研究通过运用最新开发的 RNA 靶向 CRISPR 系统 CasRx 特异性地在视网膜穆勒胶质细胞中敲低 Ptbp1 基因的表达，首次在成体中实现了视神经节细胞的再生，并且恢复了永久性视力损伤模型小鼠的视力。同时，该研究还证明了这项技术可以非常高效且特异地将纹状体内的星形胶质细胞转分化成多巴胺神经元，并且基本消除了帕金森疾病的症状。该研究将为未来众多神经退行性疾病的治疗提供一个新的途径。

人类的神经系统包含成百上千种不同类型的神经元细胞。在成熟的神经系统中，神经元一般不会再生，一旦死亡，就是永久性的。神经元的死亡会导致不同的神经退行性疾病，常见的有阿尔兹海默症和帕金森症。此类疾病的病因尚不明确且没有根治的方法，因此对人类的健康造成巨大威胁。据统计，目前全球大约有 1 亿多的人患有神经退行性疾病，而且随着老龄化的加剧，神经退行性疾病患者数量也将逐渐增多。

在常见的神经性疾病中，视神经节细胞死亡导致的永久性失明和多巴胺神经元死亡导致的帕金森疾病是尤为特殊的两类，它们都是由于特殊类型的神经元死亡导致。我们之所以能看到外界绚烂多彩的世界，是因为我们的眼睛和大脑中存在一套完整的视觉通路，而连接眼睛和大脑的神经元就是视神经节细胞。

作为眼睛和大脑的唯一一座桥梁，视神经节细胞对外界的不良刺激非常敏感。研究发现很多眼疾都可以导致视神经节细胞的死亡，急性的如缺血性视网膜病，慢性的如青光眼。视神经节细胞一旦死亡就会导致永久性失明。据统计，仅青光眼致盲的人数在全球就超过一千万人。

帕金森疾病是一种常见的老年神经退行性疾病。它的发生是由于脑内黑质区域中一种叫做多巴胺神经元的死亡，从而导致黑质多巴胺神经元不能通过黑质-纹状体通路将多巴胺运输到大脑的另一个区域纹状体。目前，全球有将近一千万人患有此病，我国尤为严重，占了大约一半的病人。 如何在成体中再生出以上两种特异类型的神经元，一直是全世界众多科学家努力的方向。

该研究中，研究人员首先在体外细胞系中筛选了高效抑制 Ptbp1 表达的 gRNA，设计了特异性标记穆勒胶质细胞和在穆勒胶质细胞中表达 CasRx 的系统。所有元件以双质粒系统的形式被包装在 AAV 中并且通过视网膜下注射，特异性地在成年小鼠的穆勒胶质细胞中下调 Ptbp1 基因的表达。

大约一个月后，研究人员在视网膜视神经节细胞层发现了由穆勒胶质细胞转分化而来的视神经节细胞，并且转分化而来的视神经节细胞可以像正常的细胞那样对光刺激产生相应的电信号。

研究人员进一步发现，转分化而来的视神经节细胞可以通过视神经和大脑中正确的脑区建立功能性的联系，并且将视觉信号传输到大脑。在视神经节细胞损伤的小鼠模型中，研究人员发现转分化的视神经细胞可以让永久性视力损伤的小鼠重新建立对光的敏感性。

为进一步发掘 Ptbp1 介导的胶质细胞向神经元转分化的治疗潜能，研究人员证明了该策略还能特异性地将纹状体中的星形胶质细胞非常高效的转分化为多巴胺神经元，并且证明了转分化而来的多巴胺神经元能够展现出和黑质中多巴胺神经元相似的特性。

在行为学测试中，研究人员发现这些转分化而来的多巴胺神经元可以弥补黑质中缺失的多巴胺神经元的功能，从而将帕金森模型小鼠的运动障碍逆转到接近正常小鼠的水平。

需要指出的是，虽然科学家们在实验室里取得了重要进展，但是要将研究成果真正应用于人类疾病的治疗，还有很多工作要做：人类的视神经节细胞能否再生？帕金森患者是否能通过该方法被治愈？这些问题有待全世界的科研工作者共同努力去寻找答案。

（上）CasRx 通过靶向的降解 Ptbp1 mRNA 从而实现 Ptbp1 基因表达的下调。

（中）视网膜下注射 AAV-GFAP-CasRx-Ptbp1 可以特异性的将视网膜穆勒胶质细胞转分化为视神经节细胞，转分化而来视神经节细胞可以和正确的脑区建立功能性的联系，并且提高永久性视力损伤模型小鼠的视力。

（下）在纹状体中注射 AAV-GFAP-CasRx-Ptbp1 可以特异性的将星形胶质细胞转分化为多巴胺神经元，从而基本消除了帕金森疾病模型小鼠的运动症状。

RNA-editing Cas13 enzymes have taken the CRISPR world by storm. Like RNA interference, these enzymes can knock down RNA without altering the genome , but Cas13s have higher on-target specificity. New work from Konermann et al. and Yan et al. describes new Cas13d enzymes that average only kb in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are the latest tools in the transcriptome engineering toolbox.

Microbial CRISPR diversity is impressive, and researchers are just beginning to tap the wealth of CRISPR possibilities. To identify Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR repeat array near a putative effector nuclease. The Cas13d proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the Cas13 superfamily. Yan et al. proceeded to study orthologs from Eubacterium siraeum (EsCas13d) and Ruminococcus sp. (RspCas13d), while Konermann et al. characterized orthologs from “Anaerobic digester metagenome” (AdmCas13d) and Ruminococcus flavefaciens (nicknamed CasRx), as well as EsCas13d.

Like other Cas13 enzymes, the Cas13d orthologs described in these papers can independently process their own CRISPR arrays into guide RNAs. crRNA cleavage is retained in dCas13d and is thus HEPN-independent. These enzymes also do not require a protospacer flanking sequence, so you can target virtually any RNA sequence ! In bacteria, Cas13d-mediated cleavage promotes collateral cleavage of other RNAs. As with other Cas13s, this collateral cleavage does not occur when Cas13d is expressed in a mammalian system.

Since Cas13d is functionally similar to previously discovered Cas13 enzymes - what makes these orthologs so special? The first property is size - Cas13d enzymes have a median length of ~930aa - making them 17-26% smaller than other Cas13s and a whopping 33% smaller than Cas9! Their small size makes then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But these studies also identified other advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both of which are described below.

The majority of Type VI-D loci contain accessory proteins with WYL domains (named for the three conserved amino acids in the domain). Yan et al. from Arbor Biotechnologies found that RspCas13d accessory protein RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins an intriguing counterpart to anti-CRISPR proteins that negatively modulate the activity of Cas enzymes, some of which are also functional in multiple species (read Arbor Biotechnologies' press release about their Cas13d deposit here ).

Not all Cas13d proteins are functional in mammalian cells, but Konermann et al. saw great results with CasRx and AdmCas13d fused to a nuclear localization signal (NLS). In a HEK293 mCherry reporter assay, CasRx and AdmCas13d produced 92% and 87% mCherry protein knockdown measured by flow cytometry, respectively. Cas13d CRISPR array processing is robust, with CasRx and either an unprocessed or processed gRNA array (22 nt spacer with 30 nt direct repeat) mediating potent knockdown. Multiplexing from the CRISPR array yielded >90% knockdown by CasRx for each of four targets, including two mRNAs and two nuclear long non-coding RNAs.

One interesting twist to Cas13d enzymes is their cleavage pattern: EsCas13d produced very similar cleavage products even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable distance from the targeted region. Konermann et al. show that EsCas13d favors cleavage at uracils, but a more detailed exploration of this cleavage pattern is necessary.

Konermann et al. compared CasRx to multiple RNA regulating methods: small hairpin RNA interference, dCas9-mediated transcriptional inhibition (CRISPRi), and Cas13a/Cas13b RNA knockdown. CasRx was the clear winner with median knockdown of 96% compared to 65% for shRNA, 53% for CRISPRi, and 66-80% for other Cas13a and Cas13b effectors. Like previously characterized Cas13 enzymes, CasRx also displays very high on-target efficiency; where shRNA treatment produced 500-900 significant off-targets, CasRx displayed zero. Unlike Cas9, for which efficiency varies widely across guide RNAs, each guide tested with CasRx yielded >80% knockdown. It seems that CasRx may make it possible to target essentially any RNA in a cell.

Since catalytically dead dCasRx maintains its RNA-binding properties, Konermann et al. tested its ability to manipulate RNA species through exon skipping. Previous CRISPR exon-skipping approaches used two guide RNAs to remove a given exon from the genome, and showed success in models of muscular dystrophy . In this case, Konermann et al. targeted MAPT , the gene encoding dementia-associated tau, delivering dCasRx and a 3-spacer array targeting the MAPT exon 10 splice acceptor and two putative splice enhancers. After AAV-mediated delivery to iPS-derived cortical neurons, dCasRx-mediated exon skipping improved the ratio of pathogenic to non-pathogenic tau by nearly 50%, showing proof-of-concept for pre-clinical and clinical applications of dCasRx.

The identification of Type VI Cas13d enzymes is another win for bioinformatic data mining. As we continue to harness the natural diversity of CRISPR systems, only time will tell how large the genome and transcriptome engineering toolbox will be. It is, however, certain that the impact of CRISPR scientific sharing will continue to grow, and we at Addgene appreciate our depositors for making their tools available to the broader community.

References

Konermann, Silvana, et al. “Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors.” Cell (2018) pii: S0092-8674(18)30207-1. PubMed PMID: 29551272

Yan, Winston X., et al. “Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein.” Mol Cell. (2018) pii: S1097-2765(18)30173-4. PubMed PMID: 29551514

\1. Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors

\2. CRISPR genetic editing takes another big step forward, targeting RNA

\3. How Editing RNA—Not DNA—Could Cure Disease in the Future

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大类学科2区。根据中科院2021年SCI期刊分区。Journal of Gastrointestinal Oncology (《胃肠肿瘤杂志》，简称JGO杂志，杂志网站)，由AME Publishing Company出版发行。该杂志的主编是美国的Gary Yang教授。 JGO于2010年9月创刊, 现已被PubMed、Free Medical Journals、Google Scholar、Index Copernicus、WZB database等国际权威数据库收录。JGO杂志主要刊载胃肠肿瘤学最新进展的论文，为读者提供有关胃肠肿瘤诊断、预防以及临床研究的新信息，包括：胃肠肿瘤多学科治疗、肿瘤标志、肿瘤影像诊断、肿瘤生物学、肿瘤病理学、癌症化学预防以及其他相关的研究进展。