中央研究院 生物化學研究所

研究領域與專長

CRISPR-Cas9系統是一套高效率基因改造技術平台，對於基因研究應用在人類細胞、動物和植物帶來突破性大變革。此系統的重要核心Cas9核酸內切酶是利用短片段guide RNA辨識目標DNA序列，並切割DNA造成雙股斷裂。藉由 Cas9 進行的雙股DNA 切割，使基因體定㸃斷裂，進而啟動細胞修補機制。CRISPR-Cas9技術不僅能達成定㸃基因剔除, 剔入和基因修飾的目的，也開啟基因治療的契機。我們實驗室將發展更具有效率、精確和安全的CRISPR-Cas9系統，應用於人類基因組編輯。目前，我們的研究方向分為下列幾項：

1. 利用基因編輯增進人類自然殺手細胞以及改善免疫細胞療法的策略。
2. 開發免疫細胞專用的腺病毒技術以增加DNA傳送效率和基因植入效率。
3. 了解CRISPR-Cas切割基因體造成的DNA雙股斷裂和修復，進而增進基因編輯效率和精確度。

學經歷

• 2006 – 2012   博士, 微生物系, 伊利諾大學香檳分校, 美國
• 2004 – 2006   碩士, 生化系, 俄亥俄州立大學, 美國
• 1996 – 2001   學士, 生物系, 英屬哥倫比亞大學, 加拿大

• 2016 – 迄今   助研究員, 中央研究院生物化學研究所
• 2012 – 2015   博士後研究員, Department of Molecular and Cell Biology, University of California, Berkeley, USA
• 2004 – 2004   研究助理, Department of Biochemistry, The Ohio State University, USA
• 2001 – 2003   研究助理, 中央研究院化學研究所

主要著作

A robust platform for expansion and genome editing of primary human natural killer cells. 
Huang RS, Lai MC, Shih HA, Lin S* 
J Exp Med. (2021)

Humanized COVID-19 decoy antibody effectively blocks viral entry and prevents SARS-CoV-2 infection. 
Huang KY, Lin MS, Kuo TC, Chen CL, Lin CC, Chou YC, Chao TL, Pang YH, Kao HC, Huang RS, Lin S, Chang SY, Yang PC 
EMBO Mol Med. (2021)

Hsc70/Stub1 promotes the removal of individual oxidatively stressed peroxisomes. 
Chen BH, Chang YJ, Lin S, Yang WY 
Nat Commun. (2020)

A multiplexed bioluminescent reporter for sensitive and non-invasive tracking of DNA double strand break repair dynamics in vitro and in vivo. 
Chien JC, Tabet E, Pinkham K, da Hora CC, Chang JC, Lin S, Badr CE, Lai CP 
Nucleic acids research (2020)

Enhanced NK-92 Cytotoxicity by CRISPR Genome Engineering Using Cas9 Ribonucleoproteins. 
Huang RS, Shih HA, Lai MC, Chang YJ, Lin S* 
Frontiers in Immunology (2020)

Generation of knock-in primary human T cells using Cas9 ribonucleoproteins. 
Schumann K, Lin S, Boyer E, Simeonov DR, Subramaniam M, Gate RE, Haliburton GE, Ye CJ, Bluestone JA, Doudna JA & Marson A 
Proc. Natl. Acad. Sci. USA (2015)

Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. 
Lin S, Staahl B & Doudna JA 
eLIFE (2014)

High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. 
Pattanayak V, Lin S, Guilinger JP, Ma E, Doudna JA & Liu DR 
Nat. Biotechnol. (2013)

Structure of the enzyme-ACP substrate gatekeeper complex required for biotin synthesis. 
Agarwal V, Lin S, Nair S & Cronan JE 
Proc. Natl. Acad. Sci. USA (2012)

Biotin synthesis begins by hijacking the fatty acid synthetic pathway. 
Lin S, Hanson RE & Cronan JE 
Nat. Chem. Biol. (2010)

著作列表

1. Chang YJ, Kang Z, Bei J, Chou SJ, Lu MJ, Su YL, Lin SW, Wang HH, (Lin S)*, Chang CJ*  (2022-06)  International journal of molecular sciences  23(12), 6839  "Generation of TRIM28 Knockout K562 Cells by CRISPR/Cas9 Genome Editing and Characterization of TRIM28-Regulated Gene Expression in Cell Proliferation and Hemoglobin Beta Subunits."
   
2. Yang TJ, Li TN, Huang RS, Pan MY, Lin SY, (Lin S), Wu KP, Wang LH, Hsu STD*  (2022-06)  JOURNAL OF CELL BIOLOGY  221(6), e202201094  "Tumor suppressor BAP1 nuclear import is governed by transportin-1."
   
3. Lin SW, Nguyen VQ, (Lin S)*  (2022-05)  BIO-PROTOCOL  12(10), e4420  "Preparation of Cas9 Ribonucleoproteins for Genome Editing"
   
4. Lin CS, Hsu CT, Yuan YH, Zheng PX, Wu FH, Cheng QW, Wu YL, Wu TL, (Lin S), Yue JJ, Cheng YH, Lin SI, Shih MC, Sheen J, Lin YC*  (2022-04)  Plant physiology  188(4), 1917-1930  "DNA-free CRISPR-Cas9 gene editing of wild tetraploid tomato Solanum peruvianum using protoplast regeneration."
   
5. Hsu CT, Yuan YH, Lin YC, (Lin S), Cheng QW, Wu FH, Sheen J, Shih MC, Lin CS  (2021-10)  The CRISPR journal  4(5), 752-760  "Efficient and Economical Targeted Insertion in Plant Genomes via Protoplast Regeneration."
   
6. Huang RS, Lai MC, (Lin S)*  (2021-09)  Current protocols  1(9), e246  "Ex Vivo Expansion and CRISPR-Cas9 Genome Editing of Primary Human Natural Killer Cells."
   
7. Hsieh HH, Chen YA, Chang YJ, Wang HH, Yu YH, Lin SW, Huang YJ, (Lin S), Chang CJ*  (2021-06)  Journal of Inflammation-London  18(1), 22  "The functional characterization of phosphorylation of tristetraprolin at C-terminal NOT1-binding domain."
   
8. Chen CL, Chen YC, Huang WL, (Lin S), Daugelavicius R, Rapoport A, Chang CR  (2021-04)  International Journal of Molecular Sciences  22(9), 4607  "A Crucial Role of Mitochondrial Dynamics in Dehydration Resistance in Saccharomyces cerevisiae."
   
9. Huang RS, Lai MC, Shih HA, (Lin S)*  (2021-03)  J Exp Med.  218(3), e20201529  "A robust platform for expansion and genome editing of primary human natural killer cells."
   
10. Huang KY, Lin MS, Kuo TC, Chen CL, Lin CC, Chou YC, Chao TL, Pang YH, Kao HC, Huang RS, (Lin S), Chang SY, Yang PC  (2021-01)  EMBO Mol Med.  13(1), e12828  "Humanized COVID-19 decoy antibody effectively blocks viral entry and prevents SARS-CoV-2 infection."
    
11. Chen BH, Chang YJ, (Lin S), Yang WY  (2020-10)  Nature Communications  11(1), 5267  "Hsc70/Stub1 promotes the removal of individual oxidatively stressed peroxisomes."
    
12. Chien JC, Tabet E, Pinkham K, da Hora CC, Chang JC, (Lin S), Badr CE, Lai CP  (2020-09)  Nucleic acids research  48(17), e100  "A multiplexed bioluminescent reporter for sensitive and non-invasive tracking of DNA double strand break repair dynamics in vitro and in vivo."
    
13. Huang RS, Shih HA, Lai MC, Chang YJ, (Lin S)*  (2020-05)  Frontiers in Immunology  11, 1008  "Enhanced NK-92 Cytotoxicity by CRISPR Genome Engineering Using Cas9 Ribonucleoproteins"
    
14. Li M, Waheed AA, Yu J, Zeng C, Chen HY, Zheng YM, Feizpour A, Reinhard BM, Gummuluru S, (Lin S), Freed EO, Liu SL  (2019-03)  Proceedings of the National Academy of Sciences of the United States of America  116(12), 5705-5714  "TIM-mediated inhibition of HIV-1 release is antagonized by Nef but potentiated by SERINC proteins."
    
15. Huang MC, Chu IT, Wang ZF, (Lin S), Chang TC, Chen CT  (2018-09)  International journal of molecular sciences  19(9), 2678  "A G-Quadruplex Structure in the Promoter Region of CLIC4 Functions as a Regulatory Element for Gene Expression."
    
16. Lai CY, Lo IW, Hewage RT, Chen YC, Chen CT, Lee CF, (Lin S), Tang MC, Lin HC  (2017-08)  Angewandte Chemie-International Edition  56(32), 9478-9482  "Biosynthesis of Complex Indole Alkaloids: Elucidation of the Concise Pathway of Okaramines."
    
17. Schumann K, (Lin S), Boyer E, Simeonov DR, Subramaniam M, Gate RE, Haliburton GE, Ye CJ, Bluestone JA, Doudna JA, Marson A  (2015-08)  Proceedings of the National Academy of Sciences of the United States of America  112(33), 10437-10442  "Generation of knock-in primary human T cells using Cas9 ribonucleoproteins."
    
18. (Lin S), Staahl BT, Alla RK, Doudna JA  (2014-12)  eLife  3, e04766  "Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery."
    
19. Jinek M, Jiang F, Taylor DW, Sternberg SH, Kaya E, Ma E, Anders C, Hauer M, Zhou K, (Lin S), Kaplan M, Iavarone AT, Charpentier E, Nogales E, Doudna JA  (2014-03)  Science  343(6176), 1247997  "Structures of Cas9 endonucleases reveal RNA-mediated conformational activation."
    
20. Lo TW, Pickle CS, (Lin S), Ralston EJ, Gurling M, Schartner CM, Bian Q, Doudna JA, Meyer BJ  (2013-10)  Genetics  195(2), 331-348  "Precise and heritable genome editing in evolutionarily diverse nematodes using TALENs and CRISPR/Cas9 to engineer insertions and deletions."
    
21. Pattanayak V, (Lin S), Guilinger JP, Ma E, Doudna JA, Liu DR  (2013-09)  Nature biotechnology  31(9), 839-843  "High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity."
    
22. Jinek M, East A, Cheng A, (Lin S), Ma E, Doudna J  (2013-01)  eLife  2, e00471  "RNA-programmed genome editing in human cells."
    
23. Agarwal V, (Lin S), Lukk T, Nair SK, Cronan JE  (2012-10)  Proceedings of the National Academy of Sciences of the United States of America  109(43), 17406-17411  "Structure of the enzyme-acyl carrier protein (ACP) substrate gatekeeper complex required for biotin synthesis."
    
24. (Lin S), Cronan JE  (2012-10)  The Journal of biological chemistry  287(44), 37010-37020  "The BioC O-methyltransferase catalyzes methyl esterification of malonyl-acyl carrier protein, an essential step in biotin synthesis."
    
25. Cronan JE, (Lin S)  (2011-06)  Current opinion in chemical biology  15(3), 407-413  "Synthesis of the alpha,omega-dicarboxylic acid precursor of biotin by the canonical fatty acid biosynthetic pathway."
    
26. (Lin S), Hanson RE, Cronan JE  (2010-09)  Nature chemical biology  6(9), 682-688  "Biotin synthesis begins by hijacking the fatty acid synthetic pathway."