李大力(華東師範大學生命科學學院研究員)

李大力(華東師範大學生命科學學院研究員)

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2004年10月至2007年5月在美國德州農工大學健康科學中心的生物科學與技術研究所從事研究助理的工作,作為聯合培的博士生於2007年6月回國獲得湖南師範大學遺傳學博士學位。畢業後受聘於華東師範大學生命科學學院生命醫學研究所從事教學與科研工作。主要負責基因敲除轉基因小鼠動物中心的建設和G蛋白偶聯受體相關的科學研究。已發表科研論文14篇,其中有10篇發表在《PNAS》、《ONCONGENE》和《ENDOCRINOLOGY》等SCI期刊上,參編專著一部,另有3篇第一作者論文和2篇合作研究論文已投稿。主持國家自然科學基金青年基金項目一項(2009-2011),作為主要成員參與上海市科委研發公共服務平台“細胞信號網路研究技術平台”建設(2007-2010)。

基本介紹

  • 中文名:李大力
  • 外文名:Dali Li
  • 國籍:中國
  • 職業:華東師範大學生命科學學院研究員
  • 畢業院校:湖南師範大學
個人簡歷,教育經歷,個人榮譽,學術兼職,研究方向,主持課題,發表論文,

個人簡歷

李大力,華東師範大學生命科學學院研究員。
2016年10月10日,來自中國科學院、北京大學、浙江大學、上海交通大學、華東師範大學、哈爾濱工業大學、溫州醫科大學等科研院所的13位課題組負責人實名公開表示,無法重複韓春雨2016年5月2日發表在《自然-生物技術》上有關NgAgo的實驗。
發表公開聲明的13位學者分別是:北京大學生命科學學院教授魏文勝,北京大學生命科學學院研究員孫育傑,北京大學分子醫學研究所教授熊敬維,中科院動物研究所研究員王皓毅、李偉,中科院生物物理研究所研究員王曉群,中科院生物化學與細胞生物學研究所研究員李勁松,中科院上海生科院神經科學研究所研究員楊輝,浙江大學生命科學研究院教授王立銘,上海交通大學教授吳強,華東師範大學生命科學學院研究員李大力,哈爾濱工業大學教授黃志偉,溫州醫科大學教授谷峰。

教育經歷

2001.9-2007.6 博士研究生 湖南師範大學生命科學學院
2004.9-2007.6 聯合培養 美國德州農工大學健康科學中心
1997.9-2001.7 本科生 湖南師範大學生命科學學院

個人榮譽

教育部“長江學者獎勵計畫”青年學者
上海市普陀區“拔尖人才”

學術兼職


  Scientific Reports編委
Nature Biotechnology, Nature Protocols, Cell Research等雜誌審稿人

研究方向

1、動物模型構建新技術開發
基因修飾動物模型是我們的主要研究對象和研究手段,課題組重點發展、最佳化基因修飾動物模型構建技術。2013年初,我們建立了基於TALEN的基因敲除小鼠平台(Nucleic Acids Research, 2013),是世界上最早利用該技術構建基因敲除小鼠的實驗室之一。在此基礎上,利用TALEN技術構建了基因敲除大鼠的技術體系,開展以基因修飾小鼠和大鼠等兩種模式動物的基礎研究(SCI CHINA LIFE SCI,2016)。 2013年8月,課題組報導了利用CRISPR/Cas9技術構建基因修飾大鼠和小鼠模型的方法,在世界上首次實現同時構建多基因敲除大鼠(Nature Biotechnology, 2013),並應邀撰寫了基因修飾大鼠構建的詳細操作步驟(Nature Protocols,2014;Methods Enzymol.2014)。通過該方法將基因修飾大鼠和小鼠的構建時間縮短到5周左右,極大的提高了效率。利用重組Cas9蛋白替代mRNA進行胚胎注射,降低脫靶活性而提升了同源重組效率,實現大片段DNA的刪除和定點基因插入,豐富了基因編輯技術在發育生物學工具小鼠和大鼠構建的套用多樣性(Scientific Reports,2015)。作為合作者,我們構建的動物模型得到廣泛套用,相關論文發表在Nature Medicine (2016),Molecular Cell(2015)和Nature Communications(2015)等雜誌。到目前為止,Cas9的脫靶問題和重組效率低一直是制約該技術進一步發展和套用的主要原因,本課題組也將重點研究如何提高同源重組效率,促進技術發展,採用非天然胺基酸定點整合的方法在哺乳動物中率先實現遺傳密碼擴充,證明哺乳動物可以耐受遺傳密碼的人為改變(Cell Research,2017)。
2、基因編輯技術在基因治療中的套用
遺傳疾病的治療往往需要長期用藥,很難達到根治的目的,極大的影響患者的生活質量,給病人和家庭帶來沉重負擔。基因治療有望在細胞中重新表達受損基因而獲得長期療效,但在實際操作中受到非常多的限制。基因編輯技術出現後,能夠對基因組DNA進行精確操作,極大降低了隨機整合的風險,有望修復基因實現長期表達,為基因治療的相關研究注入了新的活力。課題組利用CRISPR/Cas9基因編輯技術模擬在病人中新發現的凝血因子9突變,構建點突變小鼠,建立了新的B型血友病動物模型。利用定點修復成年B型血友病小鼠模型中F9基因突變策略,開展緩解凝血功能障礙的研究。通過尾靜脈注射將Cas9/sgRNA及對應的同源重組修復模板導入成年突變小鼠體內,成功地在0.56-2.84%的肝細胞中完全修復F9基因突變,顯著縮短活化部分凝血活酶時間(aPTT),極大提高了斷尾失血實驗中血友病小鼠的存活率。在國際上率先證明了通過Cas9基因編輯技術原位修復F9基因突變治療B型血友病的可行性,也為基因編輯技術套用於單基因遺傳病的治療提供了有力的實驗支持(EMBO Molecular Medicine,2016)。課題組將在此基礎上,利用AAV病毒體系導入Cas9體系,嘗試通過基因編輯治療其他遺傳疾病的可行性;通過增強基因導入效率、提高重組修復等方法獲得更高效的基因修復效率,達到更好的治療效果;建立造血幹細胞等可移植細胞基因編輯技術體系,開展離體基因編輯與細胞治療的套用研究。
3.成體幹細胞自我更新和分化機制研究
腸上皮組織是動物體中更新最快的組織之一,是理想的研究成體幹細胞分化以及組織損傷修復的模型。同時,由於多種原因導致的腸上皮的損傷也與腸炎有著密切的聯繫。腸幹細胞如何受到外界刺激引起內在信號轉導通路的變化最終表現為其自我更新或者終末分化,這是一個非常複雜而又尚待解決的重要科學問題。通過研究Lgr4/Gpr48基因敲除小鼠小腸上皮細胞分化與修復的缺陷,證明了Lgr4介導的R-spondin信號通過直接調節Wnt/beta-Catenin信號通路在腸幹細胞穩態維持和再生中的重要功能(JBC, 2013, paper of the week)。以此研究為契機迅速建立了腸幹細胞離體3D培養體系(organoid culture)和檢測方法,並利用該體外培養模型,篩選可能調節腸幹細胞增殖、分化的胞外信號分子。通過基因編輯技術,建立負責基因修飾小鼠模型,研究相關信號通路在腸幹細胞自我更新和分化的分子機制和生理病理表型。

主持課題

  1. 青春發育關鍵基因KiSS1表達調控研究(國家自然科學基金No.30800627 2009-2011 )
  2. 核仁蛋白PAK1IP1 調節核糖體生物合成的功能及機理研究 (國家自然科學基金 No.31171318 2012-2015)
  3. 七次跨膜受體Lgr4 及其配體家族在卵巢功能維持和雌性不育中的功能研究 (國家自然科學基金No.31371455 2014-2017 )
  4. 構建PAH基因新突變小鼠模型開展苯丙酮尿症基因治療研究 (國家自然科學基金 No. 81670470 2017-2020)
  5. 重症免疫缺陷型大鼠品系的構建以及基因編輯技術的最佳化研究 (上海市創新課題 No.14140900300, 2014-2017)

發表論文

5篇代表論文
  1. Li D*, Qiu Z, Shao Y, Chen Y, Guan Y, Liu M, Li Y, Gao N, Wang L, Lu X, Zhao Y*, Liu M*. Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.Nature Biotechnology. 2013 ;31(8):681-3
  2. Guan Y, Ma L, Li Q, Sun Z, Ma Li, Wu L, Wang L, Zeng L, Shao Y, Chen Y, Ma N, Lu W, Hu K, Han H, Yu Y, Huang Y, Liu M*,Li D*. CRISPR/Cas9 mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.EMBO Mol Med. 2016 May 2;8(5):477-88News&Views,Nguyen TH, Anegon I. Successful correction of hemophilia by CRISPR/Cas9 genome editing in vivo: delivery vector and immune responses are the key to success.EMBO Mol Med. 2016 May 2;8(5):439-41.) (雜誌同期點評)
  3. Chen Y, Ma J, Lu W, Tian M, Thauvin M, Yuan C, Volovitch M, Wang Q, Holst J, Liu M, Vriz S, Ye S*, Wang L*,Li D*. Heritable expansion of the genetic code in mouse and zebrafish.Cell Res. 2016 Dec 9.Editor’s Choice, Valda Vinson, Expanding the genetic code in vertebrates,Science2017:Vol. 355, Issue 6320, pp. 36 (Science雜誌編輯選為合成生物學方面當月亮點工作)
  4. Shao Y, Guan Y, Wang L, Qiu Z, Liu M, Chen Y, Wu L, Li Y, Ma X, Liu M*,Li D*. Genome editing in the rat using CRISPR-Cas and injection of RNAs into one-cell embryos.Nature Protocols. 2014 (10):2493-512
  5. Qiu Z, Liu M, Chen Z, Shao Y, Pan H, Wei G, Yu C, Zhang L, Li X, Wang P, Fan HY, Du B, Liu B, Liu M*,Li D*. High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.Nucleic Acids Res.2013 Jun;41(11):e120
論文列表:
2017:
  1. Zhang L, Li L, Gao G, Wei G, Zheng Y, Wang C, Gao N, Zhao Y, Deng J, Chen H, Sun J,Li D*, Zhang X*, Liu M*. Elevation of GPRC5A expression in colorectal cancer promotes tumor progression through VNN-1 induced oxidative stress.Int J Cancer.2017 Mar 17. doi: 10.1002/ijc.30698.
  2. Lu J, Shao Y, Qin X, Liu D, Chen A,Li D*, Liu M*, Wang X*. CRISPR knockout rat cytochrome P450 3A1/2 model for advancing drug metabolism and pharmacokinetics research.Sci Rep.2017 Feb 20;7:42922.
  3. Chen Y, Ma J, Lu W, Tian M, Thauvin M, Yuan C, Volovitch M, Wang Q, Holst J, Liu M, Vriz S, Ye S*, Wang L*,Li D*. Heritable expansion of the genetic code in mouse and zebrafish.Cell Res. 2017 Feb;27(2):294-297.
  4. Chen R, Zhang Q, Duan X, York P, Chen GD, Yin P, Zhu H, Xu M, Chen P, Wu Q, Li D, Samarut J, Xu G, Zhang P, Cao X, Li J, Wong J. The 5-hydroxymethylcytosine (5hmC) reader Uhrf2 is required for normal levels of 5hmC in mouse adult brain and spatial learning and memory.J Biol Chem.2017 Jan 23.
2016:
  1. Wei Y, Chen Y, Qiu Y, Zhao H, Liu G, Zhang Y, Meng Q, Wu G, Chen Y, Cai X, Wang H, Ying H, Zhou B, Liu M,Li D, Ding Q. Prevention of Muscle Wasting by CRISPR/Cas9-mediated Disruption of Myostatin In Vivo.Mol Ther.2016 Nov;24(11):1889-1891.
  2. You P, Hu H, Chen Y, Zhao Y, Yang Y, Wang T, Xing R, Shao Y, Zhang W,Li D*, Chen H*, Liu M*. Effects of Melanocortin 3 and 4 Receptor Deficiency on Energy Homeostasis in Rats.Sci Rep.2016 Oct 7;6:34938.
  3. Chen Y, Liu X, Zhang Y, Wang H, Ying H, Liu M,Li D, Lui KO, Ding Q. A Self-restricted CRISPR System to Reduce Off-target Effects.Mol Ther. 2016 Sep;24(9):1508-10.
  4. Zhang L, Shao Y, Li L, Tian F, Cen J, Chen X, Hu D, Zhou Y, Xie W, Zheng Y, Ji Y, Liu M,Li D*, Hui L*. Efficient liver repopulation of transplanted hepatocyte prevents cirrhosis in a rat model of hereditary tyrosinemia type I.Sci Rep. 2016 Aug 11;6:31460
  5. Chen Y, Lu W, Gao N, Long Y, Shao Y, Liu MZ, Chen H, Ye S, Ma X, Liu M*,Li D*.Generation of obese rat model by transcription activator-like effector nucleases targeting the leptin receptor gene.Sci China Life Sci.2016 May 25.
  6. Luo J, Yang Z, Ma Y, Yue Z, Lin H, Qu G, Huang J, Dai W, Li C, Zheng C, Xu L, Chen H, Wang J,Li D, Siwko S, Penninger JM, Ning G, Xiao J, Liu M. LGR4 is a receptor for RANKL and negatively regulates osteoclast differentiation and bone resorption.Nat Med. 2016 May;22(5):539-46
  7. Zhang Y, Zeng Z, Zhao J, Li D, Liu M, Wang X. Measurement of Rhodamine 123 in Three-Dimensional Organoids: A Novel Model for P-Glycoprotein Inhibitor Screening.Basic Clin Pharmacol Toxicol. 2016 Apr 6.
  8. Guan Y, Ma L, Li Q, Sun Z, Ma Li, Wu L, Wang L, Zeng L, Shao Y, Chen Y, Ma N, Lu W, Hu K, Han H, Yu Y, Huang Y, Liu M*,Li D*.CRISPR/Cas9 mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse.EMBO MOL MED. 2016 May 2;8(5):477-88.
  9. Wang X*, Tang Y, Lu J, Shao Y, Qin X, Li Y, Wang L,Li D*,Liu M*. Characterization of novel cytochrome P450 2E1 knockout rat model generated by CRISPR/Cas9.Biochem Pharmacol. 2016 Apr 1;105:80-90.
  10. Liu XM, Zhang YP, Ji SY, Li BT, Tian X, Li D, Tong C, Fan HY. Mitoguardin-1 and -2 promote maturation and the developmental potential of mouse oocytes by maintaining mitochondrial dynamics and functions.Oncotarget. 2016 Jan 12;7(2):1155-67.
2015:
  1. Wang L, Shao Y, Guan Y, Li L, Wu L, Chen F, Liu M, Chen H, Ma Y, Ma X, Liu M*,Li D*. Large genomic fragment deletion and functional gene cassette knock-in via Cas9 protein mediated genome editing in one-cell rodent embryos.Sci Rep. 2015 Dec 1;5:17517
  2. Bai M, Li Q, Shao Y, Huang Y,Li D*, Ma Y*. Generation of site-specific mutant mice using the CRISPR/Cas9 system.Yi Chuan. 2015 Oct;37(10):1029-35.
  3. Guan Y, Zhang L, Li X, Zhang X, Liu S, Gao N, Li L, Gao G, Wei G, Chen Z, Zheng Y, Ma X, Siwko S, Chen JL, Liu M*,Li D*. Repression of Mammalian Target of Rapamycin Complex 1 Inhibits Intestinal Regeneration in Acute Inflammatory Bowel Disease Models.J Immunol. 2015 May 29. pii: 1303356
  4. Liu XM, Zhang YP, Ji SY, Li BT, Tian X,Li D, Tong C, Fan HY. Mitoguardin-1 and -2 promote maturation and the developmental potential of mouse oocytes by maintaining mitochondrial dynamics and functions.Oncotarget. 2015 doi: 10.18632/oncotarget.6713
  5. Deng L, Jiang C, Chen L, Jin J, Wei J, Zhao L, Chen M, Pan W, Xu Y, Chu H, Wang X, Ge X,Li D, Liao L, Liu M, Li L, Wang P. The Ubiquitination of RagA GTPase by RNF152 Negatively Regulates mTORC1 Activation.Mol Cell. 2015 Jun 4;58(5):804-18
  6. Fan G, Sun L, Shan P, Zhang X, Huan J, Zhang X,Li D, Wang T, Wei T, Zhang X, Gu X, Yao L, Xuan Y, Hou Z, Cui Y, Cao L, Li X, Zhang S, and Wang C. Loss of KLF14 triggers centrosome amplification and tumorigenesis.Nat Commun2015 Oct 6;6:8450
  7. Fang P, Xu W,Li D, Zhao X, Dai J, Wang Z, Yan X, Qin M, Zhang Y, Xu C, Wang L, Qiao Z. A novel acrosomal protein, IQCF1, involved in sperm capacitation and the acrosome reaction.Andrology. 2015 Mar;3(2):332-44.
2014:
  1. Guan Y, Shao Y,Li D*,Liu M*. Generation of Site-Specific Mutations in the Rat Genome Via CRISPR/Cas9.Methods Enzymol.2014 ;546:297-317
  2. Shao Y, Guan Y, Wang L, Qiu Z, Liu M, Chen Y, Wu L, Li Y, Ma X, Liu M*,Li D*. Genome editing in the rat using CRISPR-Cas and injection of RNAs into one-cell embryos.Nature Protocols. 2014 (10):2493-512 (IF,9.67)
  3. Cui H, Wang Y, Huang H, Yu W, Bai M, Zhang L, Bryan BA, Wang Y, Luo J,Li D*, Ma Y*, Liu M*. GPR126 Regulates Developmental and Pathological Angiogenesis through Modulation of VEGFR2 Signaling.J Biol Chem.2014 Sep 12. pii: jbc.M114.571000 (IF,4.57)
  4. Guan X, Duan Y, Zeng Q, Pan H, Qian Y,Li D*, Cao X*, Liu M*. Lgr4 Deficiency Induces Ataxia-like Phenotype in Mice and Impairs Long-term Depression at Cerebellar Parallel fiber-Purkinje Cell Synapses.J Biol Chem.2014 Sep 19;289(38):26492-504. (IF,4.57)
  5. Pan H, Cui H, Liu S, Qian Y, Wu H, Li L, Guan Y, Guan X, Zhang L, Fan HY, Ma Y, Li R, Liu M*,Li D*. Lgr4 gene regulates Corpus Luteum Maturation through Modulation of the WNT mediated EGFR-ERK Signaling Pathway.Endocrinology. 2014 May 30:en20132183. (IF,4.50)
  6. Yi T, Weng J, Siwko S, Luo J,Li D, Liu M. LGR4/GPR48 inactivation leads to aniridia-genitourinary anomalies-mental retardation syndrome defects.J Biol Chem.2014 Mar 28;289(13):8767-80
2013:
  1. Li D*, Qiu Z, Shao Y, Chen Y, Guan Y, Liu M, Li Y, Gao N, Wang L, Lu X, Zhao Y*, Liu M*. Heritable gene targeting in the mouse and rat using a CRISPR-Cas system.Nature Biotechnol.2013 ;31(8):681-3. (IF,41.5)
  2. Qiu Z, Liu M, Chen Z, Shao Y, Pan H, Wei G, Yu C, Zhang L, Li X, Wang P, Fan HY, Du B, Liu B, Liu M,Li D*. High-efficiency and heritable gene targeting in mouse by transcription activator-like effector nucleases.Nucleic Acids Res.2013 Jun;41(11):e120. (IF,9.11)
  3. Liu S, Qian Y, Li L, Wei G, Guan Y, Pan H, Guan X, Zhang L, Lu X, Zhao Y, Liu M*,Li D*.Lgr4 deficiency increases susceptibility and severity of dextran sodium sulphate induced inflammatory bowel disease in mice.J Biol Chem. 2013 288: 8794-8803. [paper of the week] (IF,4.57)
  4. Qian Y, Liu S, Guan Y, Pan H, Guan X, Qiu Z, Li L, Gao N, Zhao Y, Li X, Lu Y, Liu M*,Li D.* Lgr4 mediated Wnt/β-catenin signaling in peritubular myoid cells is essential for spermatogenesis.Development.2013;140(8):1751-61. (IF,6.46)
  5. Luo W, Rodriguez M, Valdez JM, Zhu X, Tan K,Li D, Siwko S, Xin L, Liu M. Lgr4 is a Key Regulator of Prostate Development and Prostate Stem Cell Differentiation.Stem Cells.2013;31(11):2492-505. (IF,6.52)
  6. Wang Y, Dong J,Li D, Lai L, Siwko S, Li Y, Liu M. Lgr4 regulates mammary gland development and stem cell activity through the pluripotency transcription factor Sox2.Stem Cells. 2013;31(9):1921-31. (IF,6.52)
  7. Du B, Luo W, Li R, Tan B, Han H, Lu X,Li D, Qian M, Zhang D, Zhao Y, Liu M. Lgr4/Gpr48 negatively regulates TLR2/4-associated pattern recognition and innate immunity by targeting CD14 expression.J Biol Chem.2013; 288(21):15131-41. (IF,4.57)
2013年以前:
  1. Yu W, Qiu Z, Gao N, Wang L, Cui H, Qian Y, Jiang L, Luo J, Yi Z, Lu H,Li D*, and Liu M* PAK1IP1, a ribosomal stress-induced nucleolar protein, regulates cell proliferation via the p53-MDM2 loop,Nucleic Acids Res2010 39(6):2234-48. (IF,9.11)
  2. Yang Z, Li C, Wang X, Zhai C, Yi Z, Wang L, Liu B, Du B, Wu H, Guo X, Liu M,Li D*, Luo J*. Dauricine induces apoptosis, inhibits proliferation and invasion through inhibiting NF-kappaB signaling pathway in colon cancer cells.J Cell Physiol. 2010 Oct;225(1):266-75. (IF,3.84)
  3. Li, D., Yu, W. and Liu, M. (2009) Regulation of KiSS1 gene expression.Peptides, 30, 130-138. (IF,2.62)
  4. Li, D*., Niu, Z., Yu, W., Qian, Y., Wang, Q., Li, Q., Yi, Z., Luo, J., Wu, X., Wang, Y. Schwartz RJ, Liu M. (2009) SMYD1, the myogenic activator, is a direct target of serum response factor and myogenin.Nucleic Acids Res, 37, 7059-7071 . (IF,9.11)
  5. Li, D., Mitchell, D., Luo, J., Yi, Z., Cho, S.G., Guo, J., Li, X., Ning, G., Wu, X. and Liu, M. (2007) Estrogen regulates KiSS1 gene expression through estrogen receptor alpha and SP protein complexes.Endocrinology,148, 4821-4828. (IF,4.50)
  6. Cho SG,Li D, Tan K, Siwko SK, Liu M. KiSS1 and its G-protein-coupled receptor GPR54 in cancer development and metastasis.Cancer Metastasis Rev. 2012;31(3-4):585-91. (IF,7.24)
  7. Dai F, Chen Y, Song Y, Huang L, Zhai D, Dong Y, Lai L, Zhang T,Li D, Pang X, Liu M, Yi Z. A natural small molecule harmine inhibits angiogenesis and suppresses tumour growth through activation of p53 in endothelial cells.PLoS One. 2012;7(12):e52162 (IF,3.23)
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