請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66766完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 黃慶璨(Ching-Tsan Huang) | |
| dc.contributor.author | Stephen Kevin Chiu | en |
| dc.contributor.author | 邱煒倫 | zh_TW |
| dc.date.accessioned | 2021-06-17T00:56:29Z | - |
| dc.date.available | 2021-02-10 | |
| dc.date.copyright | 2020-02-10 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-02-04 | |
| dc.identifier.citation | Gomes AR, Byregowda SM, Veeregowda BM, Balamurugan V: An overview of heterologous expression host systems for the Production of Recombinant Proteins. Advances in Animal and Veterinary Sciences 2016, 4(7):346-356.
Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM: Heterologous protein production using the Pichia pastoris expression system. Yeast 2005, 22(4):249-270. Zahrl RJ, Pena DA, Mattanovich D, Gasser B: Systems biotechnology for protein production in Pichia pastoris. FEMS Yeast Research, 2017, 17(7). Guilliermond A: Zygosaccharomyces pastori, nouvelle espèce de levures copulation hétérogamique. Bulletin de la Société Mycologique de France 1920, 36:203-211. Phaff HJ, Miller MW, Shifrine M: The taxonomy of yeasts isolated from Drosophila in the Yosemite region of California. Antonie van Leeuwenhoek 1956, 22(1):145-161. Cregg JM, Barringer KJ, Hessler AY, Madden KR: Pichia pastoris as a host system for transformations. Molecular and Cellular Biology 1985, 5(12):3376-3385. Yamada Y, Maeda K, Mikata K: The phylogenetic relationships of the hat-shaped ascospore-forming, Nitrate-assimilating Pichia Species, Formerly Classified in the Genus Hansenula Sydow et Sydow, Based on the Partial Sequences of 18S and 26S Ribosomal RNAs (Saccharomycetaceae): The Proposals of Three New Genera, Ogataea, Kuraishia, and Nakazawaea. Bioscience, Biotechnology, and Biochemistry 1994, 58(7):1245-1257. Yamada Y, Matsuda M, Maeda K, Mikata K: The phylogenetic relationships of methanol-assimilating yeasts based on the partial Sequences of 18S and 26S Ribosomal RNAs: The Proposal of Komagataella Gen. Nov. (Saccharomycetaceae). Bioscience, Biotechnology, and Biochemistry 1995, 59(3):439-444. Kurtzman CP: Description of Komagataella phaffii sp. nov. and the transfer of Pichia pseudopastoris to the methylotrophic yeast genus Komagataella. International Journal of Systematic and Evolutionary Microbiology 2005, 55(2):973-976. Kurtzman CP: Biotechnological strains of Komagataella (Pichia) pastoris are Komagataella phaffii as determined from multigene sequence analysis. Journal of Industrial Microbiology & Biotechnology 2009, 36(11):1435-1438. Waterham HR, Russell KA, Vries Yd, Cregg JM: Peroxisomal targeting, import, and assembly of Alcohol Oxidase in Pichia pastoris. The Journal of Cell Biology 1997, 139(6):1419-1431. James M. Cregg TS: Recent Advances in the Expression of Foreign Genes in Pichia pastoris. Nature Biotechnology 1993, 11:905–910. Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM: Heterologous protein production using the Pichia pastoris expression system. Yeast 2005, 22(4):249-270. Jenkins N PR, James DC.: Getting the glycosylation right: implications for the biotechnology industry. Nature Biotechnology 1996, 14:975-981. Tokmakov AA, Kurotani A, Takagi T, Toyama M, Shirouzu M, Fukami Y, Yokoyama S: Multiple post-translational modifications affect heterologous protein synthesis. The Journal of Biological Chemistry 2012, 287(32):27106-27116. Scorer CA BR, Clare JJ, Romanos MA.: The intracellular production and secretion of HIV-1 envelope protein in the methylotrophic yeast Pichia pastoris. Gene 1993, 136(1-2):111-119. Koti Sreekrishna RGB, Keith E. Kropp, Dale T. Blankenship, Jiu-Tsair Tsay, Phil L. Smith, Jonathan D. Wierschke, Arun Subramaniam, Lori A. Birkenberger: Strategies for optimal synthesis and secretion of heterologous proteins in the methylotrophic yeast Pichia pastoris. Gene 1997, 190(1):55-62. Barrero JJ, Casler JC, Valero F, Ferrer P, Glick BS: An improved secretion signal enhances the secretion of model proteins from Pichia pastoris. Microbial Cell Factories 2018, 17(1):161. Cheng AA, Lu TK: Synthetic biology: an emerging engineering discipline. Annual Review of Biomedical Engineering 2012, 14:155-178. Liu Z, Zhang Y, Nielsen J: Synthetic Biology of Yeast. Biochemistry 2019, 58(11):1511-1520. Polizzi KM, Freemont PS: Synthetic biology biosensors for healthcare and industrial biotechnology applications. In: IET/SynbiCITE Engineering Biology Conference: 13-15 Dec. 2016 2016. 1-1. Kim HJ, Jeong H, Lee SJ: Synthetic biology for microbial heavy metal biosensors. Analytical and Bioanalytical Chemistry 2018, 410(4):1191-1203. Schwarzhans JP, Luttermann T, Geier M, Kalinowski J, Friehs K: Towards systems metabolic engineering in Pichia pastoris. Biotechnology Advances 2017, 35(6):681-710. Chang CH, Hsiung HA, Hong KL, Huang CT: Enhancing the efficiency of the Pichia pastoris AOX1 promoter via the synthetic positive feedback circuit of transcription factor Mxr1. BMC Biotechnology 2018, 18(1):81. Prabhu AA, Veeranki VD: Metabolic engineering of Pichia pastoris GS115 for enhanced pentose phosphate pathway (PPP) flux toward recombinant human interferon gamma (hIFN-gamma) production. Molecular Biology Reports 2018, 45(5):961-972. Ata O, Prielhofer R, Gasser B, Mattanovich D, Calik P: Transcriptional engineering of the glyceraldehyde-3-phosphate dehydrogenase promoter for improved heterologous protein production in Pichia pastoris. Biotechnology & Bioengineering 2017, 114(10):2319-2327. Vogl T, Kickenweiz T, Pitzer J, Sturmberger L, Weninger A, Biggs BW, Kohler EM, Baumschlager A, Fischer JE, Hyden P et al: Engineered bidirectional promoters enable rapid multi-gene co-expression optimization. Nature Communications 2018, 9(1):3589. Weis R, Luiten R, Skranc W, Schwab H, Wubbolts M, Glieder A: Reliable high-throughput screening with Pichia pastoris by limiting yeast cell death phenomena. FEMS Yeast Research 2004, 5(2):179-189. Sasagawa T, Matsui M, Kobayashi Y, Otagiri M, Moriya S, Sakamoto Y, Ito Y, Lee CC, Kitamoto K, Arioka M: High-throughput recombinant gene expression systems in Pichia pastoris using newly developed plasmid vectors. Plasmid 2011, 65(1):65-69. Kenzom T, Srivastava P, Mishra S: Simplified high-throughput screening of AOX1-expressed laccase enzyme in Pichia pastoris. Analytical Biochemistry 2015, 489:59-61. Royle KE, Polizzi K: A streamlined cloning workflow minimising the time-to-strain pipeline for Pichia pastoris. Scientific Reports 2017, 7(1):15817. Walker RSK, Pretorius IS: Applications of yeast synthetic biology geared towards the production of biopharmaceuticals. Genes (Basel) 2018, 9(7). Gasser B, Mattanovich D: A yeast for all seasons - Is Pichia pastoris a suitable chassis organism for future bioproduction? FEMS Microbiology Letters 2018, 365(17). Adli M: The CRISPR tool kit for genome editing and beyond. Nature Communications 2018, 9(1):1911. Lino CA, Harper JC, Carney JP, Timlin JA: Delivering CRISPR: a review of the challenges and approaches. Drug Delivery, 2018, 25(1):1234-1257. Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A: Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. Journal of Bacteriology 1987, 169(12):5429-5433. Mojica FJ, Diez-Villasenor C, Garcia-Martinez J, Soria E: Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. Journal of Molecular Evolution, 2005, 60(2):174-182. Martin Jinek KC, Ines Fonfara, Michael Hauer, Jennifer A. Doudna, Emmanuelle Charpentier: A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 17 Aug 2012, 337(6096). Gasiunas G, Barrangou R, Horvath P, Siksnys V: Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proceedings of the National Academy of Sciences of the United States of America 2012, 109(39):E2579-2586. Thurtle-Schmidt DM, Lo TW: Molecular biology at the cutting edge: A review on CRISPR/CAS9 gene editing for undergraduates. Biochemistry and Molecular Biology Education 2018, 46(2):195-205. Bin Moon S, Lee JM, Kang JG, Lee NE, Ha DI, Kim DY, Kim SH, Yoo K, Kim D, Ko JH et al: Highly efficient genome editing by CRISPR-Cpf1 using CRISPR RNA with a uridinylate-rich 3'-overhang. Nature Communications 2018, 9(1):3651. Eid A, Alshareef S, Mahfouz MM: CRISPR base editors: genome editing without double-stranded breaks. Biochemical Journal 2018, 475(11):1955-1964. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC et al: Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell 2014, 159(3):647-661. Dai WJ, Zhu LY, Yan ZY, Xu Y, Wang QL, Lu XJ: CRISPR-Cas9 for in vivo Gene Therapy: Promise and Hurdles.Molecular Therapy - Nucleic Acids 2016, 5:e349. Weninger A, Hatzl AM, Schmid C, Vogl T, Glieder A: Combinatorial optimization of CRISPR/Cas9 expression enables precision genome engineering in the methylotrophic yeast Pichia pastoris. Journal of Biotechnology 2016, 235:139-149. Weninger A, Fischer JE, Raschmanova H, Kniely C, Vogl T, Glieder A: Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers. Journal of Cellular Biochemistry 2018, 119(4):3183-3198. Sander JD, Joung JK: CRISPR-Cas systems for editing, regulating and targeting genomes. Nature Biotechnology 2014, 32(4):347-355. Krappmann S: CRISPR-Cas9, the new kid on the block of fungal molecular biology. Medical Mycology 2017, 55(1):16-23. Doudna FJaJA: CRISPR–Cas9 Structures and Mechanisms. Annual Review of Biophysics 2017, 46. Eoh J, Gu L: Biomaterials as vectors for the delivery of CRISPR–Cas9. Biomaterials Science 2019, 7(4):1240-1261. Liu C, Zhang L, Liu H, Cheng K: Delivery strategies of the CRISPR-Cas9 gene-editing system for therapeutic applications. Journal of Controlled Release 2017, 266:17-26. Cebrian-Serrano A, Davies B: CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools. Mammalian Genome 2017, 28(7):247-261. Moreno-Mateos MA, Vejnar CE, Beaudoin J-D, Fernandez JP, Mis EK, Khokha MK, Giraldez AJ: CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Nature Methods 2015, 12:982. Chari R, Mali P, Moosburner M, Church GM: Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach. Nature Methods 2015, 12:823. Ren X, Yang Z, Xu J, Sun J, Mao D, Hu Y, Yang S-J, Qiao H-H, Wang X, Hu Q et al: Enhanced specificity and efficiency of the CRISPR/Cas9 System with optimized sgRNA parameters in Drosophila. Cell Reports 2014, 9(3):1151-1162. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelsen TS, Heckl D, Ebert BL, Root DE, Doench JG et al: Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells. Science 2014, 343(6166):84. Wong N, Liu W, Wang X: WU-CRISPR: characteristics of functional guide RNAs for the CRISPR/Cas9 system. Genome Biology 2015, 16(1):218. Liu X, Homma A, Sayadi J, Yang S, Ohashi J, Takumi T: Sequence features associated with the cleavage efficiency of CRISPR/Cas9 system. Scientific Reports 2016, 6:19675. Wilson LOW, O’Brien AR, Bauer DC: The Current State and Future of CRISPR-Cas9 gRNA Design Tools. Frontiers in Pharmacology 2018, 9(749). Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, Smith I, Tothova Z, Wilen C, Orchard R et al: Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nature Biotechnology 2016, 34:184. Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O et al: DNA targeting specificity of RNA-guided Cas9 nucleases. Nature Biotechnology 2013, 31:827. Kouranova E, Forbes K, Zhao G, Warren J, Bartels A, Wu Y, Cui X: CRISPRs for optimal targeting: Delivery of CRISPR components as DNA, RNA, and Protein into cultured cells and single-cell Embryos. Human Gene Therapy 2016, 27(6):464-475. Xu X, Gao D, Wang P, Chen J, Ruan J, Xu J, Xia X: Efficient homology-directed gene editing by CRISPR/Cas9 in human stem and primary cells using tube electroporation. Scientific Reports 2018, 8(1):11649. Li X, Heyer W-D: Homologous recombination in DNA repair and DNA damage tolerance. Cell Research 2008, 18:99. Moore JK, Haber JE: Cell cycle and genetic requirements of two pathways of nonhomologous end-joining repair of double-strand breaks in Saccharomyces cerevisiae. Molecular and Cellular Biology 1996, 16(5):2164-2173. Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA: Enhanced proofreading governs CRISPR–Cas9 targeting accuracy. Nature 2017, 550:407. Vakulskas CA, Dever DP, Rettig GR, Turk R, Jacobi AM, Collingwood MA, Bode NM, McNeill MS, Yan S, Camarena J et al: A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. Nature Medicine 2018, 24(8):1216-1224. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, Gonzales APW, Li Z, Peterson RT, Yeh J-RJ et al: Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature 2015, 523:481. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z et al: Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 2018, 556:57. Li Q, Seys FM, Minton NP, Yang J, Jiang Y, Jiang W, Yang S: CRISPR–Cas9D10A nickase-assisted base editing in the solvent producer Clostridium beijerinckii. Biotechnology and Bioengineering 2019, 116(6):1475-1483. Deveau H, Barrangou R, Garneau JE, Labonté J, Fremaux C, Boyaval P, Romero DA, Horvath P, Moineau S: Phage Response to CRISPR-Encoded Resistance in Streptococcus thermophilus. Journal of Bacteriology 2008, 190(4):1390. Zhang Y, Heidrich N, Ampattu Biju J, Gunderson Carl W, Seifert HS, Schoen C, Vogel J, Sontheimer Erik J: Processing-Independent CRISPR RNAs Limit Natural Transformation in Neisseria meningitidis. Molecular Cell 2013, 50(4):488-503. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS et al: In vivo genome editing using Staphylococcus aureus Cas9. Nature 2015, 520:186. Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A et al: Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell 2015, 163(3):759-771. Shmakov S, Abudayyeh OO, Makarova KS, Wolf YI, Gootenberg JS, Semenova E, Minakhin L, Joung J, Konermann S, Severinov K et al: Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Molecular Cell 2015, 60(3):385-397. Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS et al: Multiplex gene editing by CRISPR–Cpf1 using a single crRNA array. Nature Biotechnology 2016, 35:31. Swiat MA, Dashko S, den Ridder M, Wijsman M, van der Oost J, Daran JM, Daran-Lapujade P: FnCpf1: a novel and efficient genome editing tool for Saccharomyces cerevisiae. Nucleic Acids Research 2017, 45(21):12585-12598. Qi Lei S, Larson Matthew H, Gilbert Luke A, Doudna Jennifer A, Weissman Jonathan S, Arkin Adam P, Lim Wendell A: Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell 2013, 152(5):1173-1183. Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein LR, Thakore PI, Glass KA, Ousterout DG, Leong KW et al: RNA-guided gene activation by CRISPR-Cas9–based transcription factors. Nature Methods 2013, 10:973. Black JB, Gersbach CA: Synthetic transcription factors for cell fate reprogramming. Current Opinion in Genetics & Development 2018, 52:13-21. O’Connell MR, Oakes BL, Sternberg SH, East-Seletsky A, Kaplan M, Doudna JA: Programmable RNA recognition and cleavage by CRISPR/Cas9. Nature 2014, 516:263. Dow LE, Fisher J, O'Rourke KP, Muley A, Kastenhuber ER, Livshits G, Tschaharganeh DF, Socci ND, Lowe SW: Inducible in vivo genome editing with CRISPR-Cas9. Nature Biotechnology 2015, 33:390. Polstein LR, Gersbach CA: A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nature Chemical Biology 2015, 11:198. Nakamura M, Srinivasan P, Chavez M, Carter MA, Dominguez AA, La Russa M, Lau MB, Abbott TR, Xu X, Zhao D et al: Anti-CRISPR-mediated control of gene editing and synthetic circuits in eukaryotic cells. Nature Communications 2019, 10(1):194. Thakore PI, Black JB, Hilton IB, Gersbach CA: Editing the epigenome: technologies for programmable transcription and epigenetic modulation. Nature Methods 2016, 13:127. Nelson CE, Wu Y, Gemberling MP, Oliver ML, Waller MA, Bohning JD, Robinson-Hamm JN, Bulaklak K, Castellanos Rivera RM, Collier JH et al: Long-term evaluation of AAV-CRISPR genome editing for Duchenne muscular dystrophy. Nature Medicine 2019, 25(3):427-432. Akcakaya P, Bobbin ML, Guo JA, Malagon-Lopez J, Clement K, Garcia SP, Fellows MD, Porritt MJ, Firth MA, Carreras A et al: In vivo CRISPR editing with no detectable genome-wide off-target mutations. Nature 2018, 561(7723):416-419. Gao Z, Herrera-Carrillo E, Berkhout B: RNA Polymerase II Activity of Type 3 Pol III Promoters. Molecular therapy Nucleic acids 2018, 12:135-145. Zheng X, Zheng P, Sun J, Kun Z, Ma Y: Heterologous and endogenous U6 snRNA promoters enable CRISPR/Cas9 mediated genome editing in Aspergillus niger. Fungal Biology and Biotechnology 2018, 5(1):2. Song L, Ouedraogo J-P, Kolbusz M, Nguyen TTM, Tsang A: Efficient genome editing using tRNA promoter-driven CRISPR/Cas9 gRNA in Aspergillus niger. PLOS ONE 2018, 13(8):e0202868. Brow DA, Guthrie C: Spliceosomal RNA U6 is remarkably conserved from yeast to mammals. Nature 1988, 334(6179):213-218. Giraud M-F, Naismith JH: The rhamnose pathway. Current Opinion in Structural Biology 2000, 10(6):687-696. Takagi Y, Sawada H: The metabolism of l-rhamnose in Escherichia coli: II. l-rhamnulose kinase. Biochimica et Biophysica Acta (BBA) - Specialized Section on Enzymological Subjects 1964, 92(1):18-25. Wegerer A, Sun T, Altenbuchner J: Optimization of an E. coli L-rhamnose-inducible expression vector: test of various genetic module combinations. BMC biotechnology 2008, 8:2-2. Liu B, Zhang Y, Zhang X, Yan C, Zhang Y, Xu X, Zhang W: Discovery of a rhamnose utilization pathway and rhamnose-inducible promoters in Pichia pastoris. Sci Rep 2016, 6:27352. Siewers V: An Overview on Selection Marker Genes for Transformation of Saccharomyces cerevisiae. In: Yeast Metabolic Engineering: Methods and Protocols 2014: 3-15. Pronk JT: Auxotrophic Yeast Strains in Fundamental and Applied Research. Applied and Environmental Microbiology 2002, 68(5):2095. Baker Brachmann C, Davies A, Cost GJ, Caputo E, Li J, Hieter P, Boeke JD: Designer deletion strains derived from Saccharomyces cerevisiae S288C: A useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 1998, 14(2):115-132. Du M, Battles MB, Nett JH: A color-based stable multi-copy integrant selection system for Pichia pastoris using the attenuated ADE1 and ADE2 genes as auxotrophic markers. Bioengineered bugs 2012, 3(1):32-37. Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH: A second set of loxP marker cassettes for Cre-mediated multiple gene knockouts in budding yeast. Nucleic Acids Research 2002, 30(6):e23-e23. Lin-Cereghino J, Hashimoto MD, Moy A, Castelo J, Orazem CC, Kuo P, Xiong S, Gandhi V, Hatae CT, Chan A et al: Direct selection of Pichia pastoris expression strains using new G418 resistance vectors. Yeast 2008, 25(4):293-299. Trastoy MO, Defais M, Larminat F: Resistance to the antibiotic Zeocin by stable expression of the Sh ble gene does not fully suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis 2005, 20(2):111-114. Papakonstantinou T, Harris S, Hearn MTW: Expression of GFP using Pichia pastoris vectors with zeocin or G-418 sulphate as the primary selectable marker. Yeast 2009, 26(6):311-321. Liu Q, Shi X, Song L, Liu H, Zhou X, Wang Q, Zhang Y, Cai M: CRISPR-Cas9-mediated genomic multiloci integration in Pichia pastoris. Microbial Cell Factories 2019, 18(1):144. Näätsaari L, Mistlberger B, Ruth C, Hajek T, Hartner FS, Glieder A: Deletion of the Pichia pastoris Ku70 homologue facilitates platform strain generation for gene expression and synthetic biology. PLOS ONE 2012, 7(6):e39720-e39720. Degreif D, Kremenovic M, Geiger T, Bertl A: Preloading budding yeast with all-in-one CRISPR/Cas9 vectors for easy and high-efficient genome editing. Journal of Biological Methods 2018, 5(3):e98. Ching-Hsiang Chang. (2017) Enhancement of Pichia pastoris AOX1 promoter efficiency by reprogramming the transcription factor Mxr1 Master Thesis, Department of Biochemical Science and Technology, College of Life Science National Taiwan University Meng-Hsi Tsai. (2017) Development of CRISPR/Cas9 system in Pichia pastoris using endogenous U6 promoter Master Thesis, Department of Biochemical Science and Technology, College of Life Science National Taiwan University | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/66766 | - |
| dc.description.abstract | 嗜甲醇酵母菌 Komagatealla phaffii (Pichia pastoris) 為目前被廣泛使用之重組蛋白質表現系統，可以甲醇誘導之強力啟動子 AOX1 大量表現重組蛋白質，且具有與哺乳動物較為相近之轉譯後修飾及可以釀酒酵母之外泌訊號胜肽α因子將蛋白質分泌至胞外之特性，常被使用於科學研究及工業發酵領域。
近年來隨著合成生物學之發展， 同時針對 Komagatealla phaffii 蛋白質表現系統內不同因子進行調控以及建構多個基因迴路之研究逐漸盛行，希望能以各式策略強化 K. phaffii 系統之各種優勢，使其能具備更高之應具價值。然而目前 K. phaffii 系統中已發表之合成生物學常見工具，如基因編輯系統、基因篩選標記回收系統、胞內 DNA 組裝工具及基因調控系統皆較為缺乏，妨礙此類研究的大幅進展。 本研究首先以前人於 K. phaffii 內建構之以 AOX1 啟動子表達 Cas9 及以 U6 啟動子表達 sgRNA 之 CRISPR/Cas9 工具將 K. phaffii 內生性基因 ADE2 及外源性基因 ZeoR 進行剔除，證明以該系統進行基因編輯之可行性。再以鼠李糖 (Rhamnose) 誘導之 LRA3 啟動子取代 AOX1 啟動子表達 hspCas9，避免影響 AOX1 啟動子之重組蛋白質表達量，並配合在質體上加入目標基因之同源重組模板及改變誘導表現 hspCas9 之流程，以提升 CRISPR/Cas9 基因編輯工具之效率，進一步改善本實驗室建構之 U6 CRISPR/Cas9 基因編輯工具之應用性。 | zh_TW |
| dc.description.abstract | The methylotrophic yeast Komagataella phaffii (Syn. Pichia pastoris) is a recombinant protein expression system who has a high expression level methanol inducible promoter AOX1, an post-translational modification similar to human cell in eukaryotic systems and ability to secret recombinant protein by Saccharomyces cerevisiae α-factor. These advantages make it widely used in industrial and research field.
Along with synthetic biology become popular in recent years, there is more and more research aimed to regulated different factor and establish more complicated gene circuit. Try to improving K. phaffii’s capability and value. However, because the lack of some basic synthetic biology tools, such as genome editing tools, marker recycle tools, in vivo DNA assemble tools and gene regulation system, this type of research is still obstructed. This thesis wants to continue the previous research which establishes a CRISPR/Cas9 genome editing system in K. phaffii used AOX1 promoter express hspCas9 and endogenous U6 promoter express sgRNA. First of all, we choose K. phaffii 's auxotrophy marker gene ADE2 and drug resistance mafrker gene ZeoR as a target gene to knock out by the CRISPR/Cas9 system. Then we replace the AOX1 promoter by a rhamnose-inducible promoter LRA3 to prevent this CRISPR/Cas9 system impact the recombinant protein's yield. At last, we combine two strategies, an episomal plasmid contain homologous DNA Donor and improve the induction process, together to increase the CRISPR/Cas9 genome editing efficiency. Make the K. phaffii U6 CRISPR/Cas9 genome editing tool more applicability. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T00:56:29Z (GMT). No. of bitstreams: 1 ntu-109-R06b22049-1.pdf: 33432525 bytes, checksum: 10b25c5420ce2906f8fae8641b9d9cf0 (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 第一章 前言 .............. 1
一、異源蛋白質表達系統 (heterologous protein expression system) .............. 1 1. Komagataella phaffii (Pichia pastoris) 簡介 .............. 4 2. Komagataella phaffii (Pichia pastoris) 異源蛋白質表達系統 .............. 5 3. Komagataella phaffii (Pichia pastoris)之合成生物學研究 .............. 5 二、CRISPR 基因編輯系統 .............. 6 1. CRISPR 研究之沿革 .............. 7 2. CRISPR/Cas9 基因編輯系統之作用機制 .............. 8 3. CRISPR/Cas9 基因編輯系統之運送策略 .............. 11 4. 影響 CRISPR/Cas9 基因編輯系統效率之因素 .............. 12 5. Komagataella phaffii 之 CRISPR/Cas9 基因編輯研究 .............. 15 6. CRISPR 其他應用與發展 .............. 16 三. Komagataella phaffii 之 U6 啟動子 .............. 19 四. Komagataella phaffii 之 LRA3 啟動子 .............. 20 五.基因篩選標記 .............. 21 六.研究動機 .............. 24 1.目的 .............. 25 2.提升基因編輯效率之策略 .............. 26 3.目標 .............. 26 第二章 材料方法 .............. 29 一、藥品與培養基 .............. 29 二、使用菌株與培養條件 .............. 32 三、表現載體之建構…… .............. 33 四、Komagatella phaffii 電穿孔轉形法 .............. 40 五、Komagatella phaffii 轉形株之基因分析 .............. 41 六、Komagatella phaffii 轉形株之試管培養、誘導啟動 CRISPR/Cas9 基因編輯系統 .............. 42 七、Komagatella phaffii 轉形株之搖瓶培養、誘導及分析 .............. 43 八、CRISPR/Cas9 基因編輯系統 sgRNA 之設計 .............. 43 九、Komagatella phaffii 轉形株之 U6 啟動子轉錄 sgRNA 確認 .............. 43 十、蛋白質產物分析 .............. 44 十一、CRISPR/Cas9 基因編輯系統效率計算 .............. 45 第三章 結果 .............. 47 一、 確認 Komagataella phaffii CRISPR/Cas9 基因編輯系統之功能 .............. 47 1. 表現 hspCas9 轉形株之篩選 .............. 47 2. 甲醇誘導表現 hspCas9 重組蛋白質 .............. 47 3. 同時表現 sgRNA 與 hspCas9 之轉形株篩選 .............. 48 4. 確認 sgRNA 之表現 .............. 48 5. 以甲醇誘導進行 CRISPR 基因編輯作用 .............. 48 二、以 Komagataella phaffii 內生 LRA3 啟動子重新建構 CRISPR/Cas9 基因編輯系統 .............. 58 1. 釣取 Komagataella phaffii 之 LRA3 啟動子 .............. 58 2. 表現 mEGFP 之轉形株篩選及表現 .............. 58 3. 表現 hspCas9 之轉形株篩選 .............. 59 4. 同時表現 sgRNA 與 hspCas9 之轉形株篩選 .............. 59 5. 以鼠李糖誘導進行 CRISPR 基因編輯作用 .............. 60 三、以同源重組策略提升基因編輯效率 .............. 71 1. 以帶有同源重組模板之游離質體提升基因編輯效率 .............. 71 2. 改良誘導策略提升基因編輯效率 .............. 72 四、在 K. phaffii 中以 CRISPR/Cas9 基因編輯工具建構基因篩選標記回收系統 .............. 78 1.設計篩選標記基因回收質體並建構菌株 .............. 78 2.以鼠李糖誘導回收篩選標記基因 ZeoR .............. 78 第四章 討論 .............. 84 一、以 K. phaffii 內生性基因 ADE2 與外源抗藥性基因 ZeoR 取代外源綠色螢光蛋白基因 mEGFP 做為 CRISPR/Cas9 基因編輯工具之目標 .............. 84 二、發展 K. phaffii CRISPR/Cpf1 基因編輯工具 .............. 85 三、以 LRA3 啟動子取代 AOX1 啟動子表現 hspCas9 進行基因編輯之效果 .............. 86 四、以提供同源重組模板以及同時放大培養與誘導表現 hspCas9 之策略提升基因編輯效率 .............. 87 五、本研究建構 K. phaffii CRISPR/Cas9 基因編輯工具與現存其他 K. phaffii CRISPR/Cas9 基因編輯工具之比較 .............. 88 六、 本研究以 K. phaffii CRISPR/Cas9 基因編輯工具建構之篩選標記基因回收系統之優缺點 .............. 9 第五章 結論 .............. 91 第六章 未來展望 .............. 92 一、以其他適合之啟動子取代 LRA3 啟動子 .............. 92 二、發展 K. phaffii CRISPR/Cpf1 基因編輯工具 .............. 92 三、以 K. phaffii CRISPR/Cas9 基因編輯工具於染色體特定位點插入基因序列 .............. 93 四、嘗試同時進行多基因編輯 .............. 93 五、探討表現 hspCas9 以及CRISPR/Cas9 基因編輯工具對於 K. phaffii 菌株之影響 .............. 94 六、運用 K. phaffii CRISPR/Cas9 基因編輯工具建構基因篩選標記回收系統 .............. 94 第七章 參考文獻 .............. 95 | |
| dc.language.iso | zh-TW | |
| dc.subject | 重組蛋白質 | zh_TW |
| dc.subject | 篩選標記基因回收 | zh_TW |
| dc.subject | 嗜甲醇酵母菌 | zh_TW |
| dc.subject | 基因編輯 | zh_TW |
| dc.subject | Pichia pastoris | en |
| dc.subject | CRISPR/Cas9 | en |
| dc.subject | Komagataella phaffii | en |
| dc.title | 在 Komagataella phaffii 中以同源重組策略提升
CRISPR/Cas9 基因編輯效率 | zh_TW |
| dc.title | Enhancement of CRISPR/Cas9 genome editing efficiency
using homologus recombination in Komagataella phaffii | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林晉玄(Ching-hsuan Lin),張世宗(Shih-Chung Zhang),陳浩仁(Hau-Ren Chen),凌嘉鴻 | |
| dc.subject.keyword | 嗜甲醇酵母菌,基因編輯,篩選標記基因回收,重組蛋白質, | zh_TW |
| dc.subject.keyword | CRISPR/Cas9,Pichia pastoris,Komagataella phaffii, | en |
| dc.relation.page | 105 | |
| dc.identifier.doi | 10.6342/NTU202000245 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2020-02-04 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-109-1.pdf 未授權公開取用 | 32.65 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。