Synthetic biology and viral vaccine development

doi:10.12211/2096-8280.2022-064

Abstract

Abstract:

Infectious diseases caused by viruses seriously endanger public health, and thus pose a great impact on socioeconomic development. Vaccine development is a critical and effective strategy for preventing the spread of infectious diseases to control them effectively. Generally, viral vaccines can be divided into various categories, such as whole virus vaccines (e.g. inactivated virus vaccines, split inactivated vaccines, and live attenuated vaccines), nucleic acid vaccines (DNA and RNA vaccines), recombinant subunits vaccines, and viral vector-based vaccines. However, strategies for developing viral vaccines still face some challenges, such as time-consuming, limited efficacy, and safety concern, which hinder their development, especially for fighting emerging infectious diseases timely. With the rapid development of synthetic biology, novel vaccines, named as synthetic vaccines, including genomic codon-optimized vaccines, nucleic acid vaccines, viral vector vaccines, virus-particle-like vaccines, and cell-based vaccines, have been designed, which can elicit immune protection more effectively. Synthetic biology technologies, such as codon optimization/deoptimization, genetically encoded click chemistry, and bioconjugation, can overcome weaknesses of traditional vaccines, and in the meantime facilitate the development of safe and efficient virus synthetic vaccines, which have been extensively explored. In this review, we summarize the current status of traditional vaccines, and also address the potential applications and advantages of synthetic biology technologies in the development of viral vaccines. At the end, we highlight the challenges of synthetic vaccines, which may provide insights and guidances for their design.

Key words: viral vaccines, synthetic biology, synthetic vaccines, genomic codon-optimized vaccines

摘要：

病毒所致的传染性疾病严重危害公共卫生安全，给全球人类健康与经济发展带来了巨大的威胁。疫苗是防治传染性疾病传播的一个关键且有效的手段，包括全病毒疫苗、亚单位疫苗、核酸疫苗等。然而，现有的疫苗开发策略面临研发周期、有效性、安全性等系列问题，是当今应对新发传染病的难点和痛点。利用合成生物学技术，比如密码子优化技术、可基因编码点击化学技术、生物偶联技术等，可以克服以上问题，在短时间内研发出安全且高效的病毒合成疫苗，因此合成生物学技术在病毒合成疫苗研发中的应用越来越广泛。本文总结了传统病毒疫苗的现状与应用，进而阐述了合成生物学技术在病毒疫苗研发中的应用与优势，并提出了病毒合成疫苗将要面临的挑战，为设计下一代新型病毒疫苗提供思路和方法。

关键词: 病毒疫苗, 合成生物学, 合成疫苗, 密码子优化疫苗

CLC Number:

R373

Zhaoling SHEN, Yanling WU, Tianlei YING. Synthetic biology and viral vaccine development[J]. Synthetic Biology Journal, 2023, 4(2): 333-346.

申赵铃, 吴艳玲, 应天雷. 合成生物学与病毒疫苗研发[J]. 合成生物学, 2023, 4(2): 333-346.

Figures/Tables 4

References 102

1	ROYCHOUDHURY S, DAS A, SENGUPTA P, et al. Viral pandemics of the last four decades: pathophysiology, health impacts and perspectives[J]. International Journal of Environmental Research and Public Health, 2020, 17(24): 9411.
2	AKIN L, GÖZEL M G. Understanding dynamics of pandemics[J]. Turkish Journal of Medical Sciences, 2020, 50(SI-1): 515-519.
3	MEYER H, EHMANN R, SMITH G L. Smallpox in the post-eradication era[J]. Viruses, 2020, 12(2): 138.
4	AHMED R, BURTON D R. Viral vaccines: past successes and future challenges[J]. Current Opinion in Virology, 2013, 3(3): 307-308.
5	BRISSE M, VRBA S M, KIRK N, et al. Emerging concepts and technologies in vaccine development[J]. Frontiers in Immunology, 2020, 11: 583077.
6	MENG F K, ELLIS T. The second decade of synthetic biology: 2010—2020[J]. Nature Communications, 2020, 11: 5174.
7	HO C, MORSUT L. Novel synthetic biology approaches for developmental systems[J]. Stem Cell Reports, 2021, 16(5): 1051-1064.
8	TAN X, LETENDRE J H, COLLINS J J, et al. Synthetic biology in the clinic: engineering vaccines, diagnostics, and therapeutics[J]. Cell, 2021, 184(4): 881-898.
9	WANG N X, YUAN Z, NIU W, et al. Synthetic biology approach for the development of conditionally replicating HIV-1 vaccine[J]. Journal of Chemical Technology and Biotechnology, 2017, 92(3): 455-462.
10	GHATTAS M, DWIVEDI G, LAVERTU M, et al. Vaccine technologies and platforms for infectious diseases: current progress, challenges, and opportunities[J]. Vaccines, 2021, 9(12): 1490.
11	AWADASSEID A, WU Y L, TANAKA Y, et al. Current advances in the development of SARS-CoV-2 vaccines[J]. International Journal of Biological Sciences, 2021, 17(1): 8-19.
12	ARORA M, LAKSHMI R. Vaccines-safety in pregnancy[J]. Best Practice & Research Clinical Obstetrics & Gynaecology, 2021, 76: 23-40.
13	CURRAN M P, LEROUX-ROELS I. Inactivated split-virion seasonal influenza vaccine (Fluarix): a review of its use in the prevention of seasonal influenza in adults and the elderly[J]. Drugs, 2010, 70(12): 1519-1543.
14	CHEN H P, HUANG Z Y, CHANG S Y, et al. Immunogenicity and safety of an inactivated SARS-CoV-2 vaccine (Sinopharm BBIBP-CorV) coadministered with quadrivalent split-virion inactivated influenza vaccine and 23-valent pneumococcal polysaccharide vaccine in China: a multicentre, non-inferiority, open-label, randomised, controlled, phase 4 trial[J]. Vaccine, 2022, 40(36): 5322-5332.
15	STANFIELD B A, KOUSOULAS K G, FERNANDEZ A, et al. Rational design of live-attenuated vaccines against herpes simplex viruses[J]. Viruses, 2021, 13(8): 1637.
16	MOK D Z L, CHAN K R. The effects of pre-existing antibodies on live-attenuated viral vaccines[J]. Viruses, 2020, 12(5): 520.
17	VETTER V, DENIZER G, FRIEDLAND L R, et al. Understanding modern-day vaccines: what you need to know[J]. Annals of Medicine, 2018, 50(2): 110-120.
18	MOYLE P M, TOTH I. Modern subunit vaccines: development, components, and research opportunities[J]. ChemMedChem, 2013, 8(3): 360-376.
19	HANSSON M, NYGREN P A, STÅHL S. Design and production of recombinant subunit vaccines[J]. Biotechnology and Applied Biochemistry, 2000, 32(2): 95-107.
20	ZHANG N R, ZHENG B J, LU L, et al. Advancements in the development of subunit influenza vaccines[J]. Microbes and Infection, 2015, 17(2): 123-134.
21	AZMI F, HADI AHMAD FUAAD A AL, SKWARCZYNSKI M, et al. Recent progress in adjuvant discovery for peptide-based subunit vaccines[J]. Human Vaccines & Immunotherapeutics, 2014, 10(3): 778-796.
22	MCVOY M A. Cytomegalovirus vaccines[J]. Clinical Infectious Diseases, 2013, 57(): S196-S199.
23	PASS R F, ZHANG C P, EVANS A, et al. Vaccine prevention of maternal cytomegalovirus infection[J]. The New England Journal of Medicine, 2009, 360(12): 1191-1199.
24	TOMBÁCZ I, WEISSMAN D, PARDI N. Vaccination with messenger RNA: a promising alternative to DNA vaccination[J]. Methods in Molecular Biology, 2021, 2197: 13-31.
25	GRANT-KLEIN R J, ALTAMURA L A, BADGER C V, et al. Codon-optimized filovirus DNA vaccines delivered by intramuscular electroporation protect cynomolgus macaques from lethal Ebola and Marburg virus challenges[J]. Human Vaccines & Immunotherapeutics, 2015, 11(8): 1991-2004.
26	LEDWITH B J, MANAM S, TROILO P J, et al. Plasmid DNA vaccines: investigation of integration into host cellular DNA following intramuscular injection in mice[J]. Intervirology, 2000, 43(4/5/6): 258-272.
27	WANG Z, TROILO P J, WANG X, et al. Detection of integration of plasmid DNA into host genomic DNA following intramuscular injection and electroporation[J]. Gene Therapy, 2004, 11(8): 711-721.
28	SZABÓ G T, MAHINY A J, VLATKOVIC I. COVID-19 mRNA vaccines: platforms and current developments[J]. Molecular Therapy, 2022, 30(5): 1850-1868.
29	KARIKÓ K. Developing mRNA for therapy[J]. The Keio Journal of Medicine, 2022, 71(1): 31.
30	LI M Y, WANG Z N, XIE C Y, et al. Advances in mRNA vaccines[J]. International Review of Cell and Molecular Biology, 2022, 372: 295-316.
31	WANG Y, ZHANG Z Q, LUO J W, et al. mRNA vaccine: a potential therapeutic strategy[J]. Molecular Cancer, 2021, 20(1): 33.
32	FANG E Y, LIU X H, LI M, et al. Advances in COVID-19 mRNA vaccine development[J]. Signal Transduction and Targeted Therapy, 2022, 7: 94.
33	MCCANN N, O'CONNOR D, LAMBE T, et al. Viral vector vaccines[J]. Current Opinion in Immunology, 2022, 77: 102210.
34	EWER K J, LAMBE T, ROLLIER C S, et al. Viral vectors as vaccine platforms: from immunogenicity to impact[J]. Current Opinion in Immunology, 2016, 41: 47-54.
35	CHEN J D, WANG J H, ZHANG J P, et al. Advances in development and application of influenza vaccines[J]. Frontiers in Immunology, 2021, 12: 711997.
36	DE VRIES R D, RIMMELZWAAN G F. Viral vector-based influenza vaccines[J]. Human Vaccines & Immunotherapeutics, 2016, 12(11): 2881-2901.
37	KHAN M S, KIM E, MCPHERSON A, et al. Adenovirus-vectored SARS-CoV-2 vaccine expressing S1-N fusion protein[J]. Antibody Therapeutics, 2022, 5(3): 177-191.
38	TOURNIER J N, KONONCHIK J. Virus eradication and synthetic biology: changes with SARS-CoV-2?[J]. Viruses, 2021, 13(4): 569.
39	PARVATHY S T, UDAYASURIYAN V, BHADANA V. Codon usage bias[J]. Molecular Biology Reports, 2022, 49(1): 539-565.
40	XU Y C, LIU K S, HAN Y, et al. Codon usage bias regulates gene expression and protein conformation in yeast expression system P. pastoris [J]. Microbial Cell Factories, 2021, 20(1): 91.
41	NOVOA E M, PAVON-ETERNOD M, PAN T, et al. A role for tRNA modifications in genome structure and codon usage[J]. Cell, 2012, 149(1): 202-213.
42	NOVOA E M, RIBAS DE POUPLANA L. Speeding with control: codon usage, tRNAs, and ribosomes[J]. Trends in Genetics, 2012, 28(11): 574-581.
43	GUIMARAES J C, MITTAL N, GNANN A, et al. A rare codon-based translational program of cell proliferation[J]. Genome Biology, 2020, 21(1): 44.
44	FU H G, LIANG Y B, ZHONG X Q, et al. Codon optimization with deep learning to enhance protein expression[J]. Scientific Reports, 2020, 10(1): 17617.
45	FENG L Q, WANG Q, SHAN C, et al. An adenovirus-vectored COVID-19 vaccine confers protection from SARS-CoV-2 challenge in rhesus macaques[J]. Nature Communications, 2020, 11: 4207.
46	XU S Q, YANG K P, LI R, et al. mRNA vaccine era-mechanisms, drug platform and clinical prospection[J]. International Journal of Molecular Sciences, 2020, 21(18): 6582.
47	COLEMAN J R, PAPAMICHAIL D, SKIENA S, et al. Virus attenuation by genome-scale changes in codon pair bias[J]. Science, 2008, 320(5884): 1784-1787.
48	BROADBENT A J, SANTOS C P, ANAFU A, et al. Evaluation of the attenuation, immunogenicity, and efficacy of a live virus vaccine generated by codon-pair bias de-optimization of the 2009 pandemic H1N1 influenza virus, in ferrets[J]. Vaccine, 2016, 34(4): 563-570.
49	KAPLAN B S, SOUZA C K, GAUGER P C, et al. Vaccination of pigs with a codon-pair bias de-optimized live attenuated influenza vaccine protects from homologous challenge[J]. Vaccine, 2018, 36(8): 1101-1107.
50	MARUGGI G, ZHANG C L, LI J W, et al. mRNA as a transformative technology for vaccine development to control infectious diseases[J]. Molecular Therapy, 2019, 27(4): 757-772.
51	BURNS C C, SHAW J, CAMPAGNOLI R, et al. Modulation of poliovirus replicative fitness in HeLa cells by deoptimization of synonymous codon usage in the capsid region[J]. Journal of Virology, 2006, 80(7): 3259-3272.
52	SI L L, XU H, ZHOU X Y, et al. Generation of influenza A viruses as live but replication-incompetent virus vaccines[J]. Science, 2016, 354(6316): 1170-1173.
53	JOHNSON B A, XIE X P, BAILEY A L, et al. Loss of furin cleavage site attenuates SARS-CoV-2 pathogenesis[J]. Nature, 2021, 591(7849): 293-299.
54	GOŁAWSKI M, LEWANDOWSKI P, JABŁOŃSKA I, et al. The reassessed potential of SARS-CoV-2 attenuation for COVID-19 vaccine development - a systematic review[J]. Viruses, 2022, 14(5): 991.
55	WANG Y, YANG C, SONG Y T, et al. Scalable live-attenuated SARS-CoV-2 vaccine candidate demonstrates preclinical safety and efficacy[J]. Proceedings of the National Academy of Sciences of the United States of America, 2021, 118(29): e2102775118.
56	FAN R L Y, VALKENBURG S A, WONG C K S, et al. Generation of live attenuated influenza virus by using codon usage bias[J]. Journal of Virology, 2015, 89(21): 10762-10773.
57	ASRANI K H, FARELLI J D, STAHLEY M R, et al. Optimization of mRNA untranslated regions for improved expression of therapeutic mRNA[J]. RNA Biology, 2018, 15(6): 756-762.
58	TREPOTEC Z, ANEJA M K, GEIGER J, et al. Maximizing the translational yield of mRNA therapeutics by minimizing 5′-UTRs[J]. Tissue Engineering Part A, 2019, 25(1/2): 69-79.
59	VON NIESSEN A G O, POLEGANOV M A, RECHNER C, et al. Improving mRNA-based therapeutic gene delivery by expression-augmenting 3′ UTRs identified by cellular library screening[J]. Molecular Therapy, 2019, 27(4): 824-836.
60	PARDI N, HOGAN M J, WEISSMAN D. Recent advances in mRNA vaccine technology[J]. Current Opinion in Immunology, 2020, 65: 14-20.
61	LEE S, RYU J H. Influenza viruses: innate immunity and mRNA vaccines[J]. Frontiers in Immunology, 2021, 12: 710647.
62	NOORI M, NEJADGHADERI S A, ARSHI S, et al. Potency of BNT162b2 and mRNA-1273 vaccine-induced neutralizing antibodies against severe acute respiratory syndrome-CoV-2 variants of concern: a systematic review of in vitro studies[J]. Reviews in Medical Virology, 2022, 32(2): e2277.
63	FEIKIN D R, HIGDON M M, ABU-RADDAD L J, et al. Duration of effectiveness of vaccines against SARS-CoV-2 infection and COVID-19 disease: results of a systematic review and meta-regression[J]. The Lancet, 2022, 399(10328): 924-944.
64	XI J X, LEI L R, ZOUZAS W, et al. Nasally inhaled therapeutics and vaccination for COVID-19: developments and challenges[J]. MedComm, 2021, 2(4): 569-586.
65	ANDRADE V M, CHRISTENSEN-QUICK A, AGNES J, et al. INO-4800 DNA vaccine induces neutralizing antibodies and T cell activity against global SARS-CoV-2 variants[J]. Npj Vaccines, 2021, 6: 121.
66	SMITH T R F, PATEL A, RAMOS S, et al. Immunogenicity of a DNA vaccine candidate for COVID-19[J]. Nature Communications, 2020, 11: 2601.
67	WALTERS J N, SCHOUEST B, PATEL A, et al. Prime-boost vaccination regimens with INO-4800 and INO-4802 augment and broaden immune responses against SARS-CoV-2 in nonhuman primates[J]. Vaccine, 2022, 40(21): 2960-2969.
68	DEY A, CHOZHAVEL RAJANATHAN T M, CHANDRA H, et al. Immunogenic potential of DNA vaccine candidate, ZyCoV-D against SARS-CoV-2 in animal models[J]. Vaccine, 2021, 39(30): 4108-4116.
69	KHOBRAGADE A, BHATE S, RAMAIAH V, et al. Efficacy, safety, and immunogenicity of the DNA SARS-CoV-2 vaccine (ZyCoV-D): the interim efficacy results of a phase 3, randomised, double-blind, placebo-controlled study in India[J]. The Lancet, 2022, 399(10332): 1313-1321.
70	ALMEIDA A M, EUSÉBIO D, QUEIROZ J A, et al. Minicircle DNA vaccine purification and E7 antigen expression assessment[J]. Methods in Molecular Biology, 2021, 2197: 207-222.
71	JIANG Y L, GAO X, XU K, et al. A novel cre recombinase-mediated in vivo minicircle DNA (CRIM) vaccine provides partial protection against Newcastle disease virus[J]. Applied and Environmental Microbiology, 2019, 85(14): e00407-e00419.
72	ZHOU X W, JIANG X, QU M Y, et al. Engineering antiviral vaccines[J]. ACS Nano, 2020, 14(10): 12370-12389.
73	TATSIS N, ERTL H C J. Adenoviruses as vaccine vectors[J]. Molecular Therapy, 2004, 10(4): 616-629.
74	SAKURAI F, TACHIBANA M, MIZUGUCHI H. Adenovirus vector-based vaccine for infectious diseases[J]. Drug Metabolism and Pharmacokinetics, 2022, 42: 100432.
75	SHIRLEY J L, DE JONG Y P, TERHORST C, et al. Immune responses to viral gene therapy vectors[J]. Molecular Therapy, 2020, 28(3): 709-722.
76	TOMORI O, KOLAWOLE M O. Ebola virus disease: current vaccine solutions[J]. Current Opinion in Immunology, 2021, 71: 27-33.
77	YAMAZAKI K I, DE MORA K, SAITOH K. BioBrick-based 'quick gene assembly' in vitro [J]. Synthetic Biology, 2017, 2(1): ysx003.
78	SHETTY R P, ENDY D, KNIGHT T F. Engineering BioBrick vectors from BioBrick parts[J]. Journal of Biological Engineering, 2008, 2: 5.
79	NOORAEI S, BAHRULOLUM H, HOSEINI Z S, et al. Virus-like particles: preparation, immunogenicity and their roles as nanovaccines and drug nanocarriers[J]. Journal of Nanobiotechnology, 2021, 19(1): 59.
80	PATEL J M, KIM M C, VARTABEDIAN V F, et al. Protein transfer-mediated surface engineering to adjuvantate virus-like nanoparticles for enhanced anti-viral immune responses[J]. Nanomedicine: Nanotechnology, Biology, and Medicine, 2015, 11(5): 1097-1107.
81	MOHSEN M O, ZHA L S, CABRAL-MIRANDA G, et al. Major findings and recent advances in virus-like particle (VLP)-based vaccines[J]. Seminars in Immunology, 2017, 34: 123-132.
82	LAMPINEN V, HEINIMÄKI S, LAITINEN O H, et al. Modular vaccine platform based on the norovirus-like particle[J]. Journal of Nanobiotechnology, 2021, 19(1): 25.
83	COHEN A A, GNANAPRAGASAM P N P, LEE Y E, et al. Mosaic nanoparticles elicit cross-reactive immune responses to zoonotic coronaviruses in mice[J]. Science, 2021, 371(6530): 735-741.
84	CHARLTON HUME H K, VIDIGAL J, CARRONDO M J T, et al. Synthetic biology for bioengineering virus-like particle vaccines[J]. Biotechnology and Bioengineering, 2019, 116(4): 919-935.
85	TEYMENNET-RAMÍREZ K V, MARTÍNEZ-MORALES F, TREJO-HERNÁNDEZ M R. Yeast surface display system: strategies for improvement and biotechnological applications[J]. Frontiers in Bioengineering and Biotechnology, 2021, 9: 794742.
86	KUMAR R, KUMAR P. Yeast-based vaccines: new perspective in vaccine development and application[J]. FEMS Yeast Research, 2019, 19(2): foz007.
87	POLLET J, CHEN W H, VERSTEEG L, et al. SARS‑CoV-2 RBD219-N1C1: a yeast-expressed SARS-CoV-2 recombinant receptor-binding domain candidate vaccine stimulates virus neutralizing antibodies and T-cell immunity in mice[J]. Human Vaccines & Immunotherapeutics, 2021, 17(8): 2356-2366.
88	LEE J, LIU Z Y, CHEN W H, et al. Process development and scale-up optimization of the SARS-CoV-2 receptor binding domain-based vaccine candidate, RBD219-N1C1[J]. Applied Microbiology and Biotechnology, 2021, 105(10): 4153-4165.
89	ZHAO Q J, TOWNE V, BROWN M, et al. In-depth process understanding of RECOMBIVAX HB^® maturation and potential epitope improvements with redox treatment: multifaceted biochemical and immunochemical characterization[J]. Vaccine, 2011, 29(45): 7936-7941.
90	WANG J W, RODEN R B. Virus-like particles for the prevention of human papillomavirus-associated malignancies[J]. Expert Review of Vaccines, 2013, 12(2): 129-141.
91	VENTER J C, GLASS J I, C A Ⅲ HUTCHISON, et al. Synthetic chromosomes, genomes, viruses, and cells[J]. Cell, 2022, 185(15): 2708-2724.
92	WANG W H, ERAZO E M, ISHCOL M R C, et al. Virus-induced pathogenesis, vaccine development, and diagnosis of novel H7N9 avian influenza A virus in humans: a systemic literature review[J]. The Journal of International Medical Research, 2020, 48(1): 300060519845488.
93	TRIPATHI N K, SHRIVASTAVA A. Recent developments in recombinant protein-based dengue vaccines[J]. Frontiers in Immunology, 2018, 9: 1919.
94	PENG X L, CHENG J S Y, GONG H L, et al. Advances in the design and development of SARS-CoV-2 vaccines[J]. Military Medical Research, 2021, 8(1): 67.
95	CALZAS C, CHEVALIER C. Innovative mucosal vaccine formulations against influenza A virus infections[J]. Frontiers in Immunology, 2019, 10: 1605.
96	NAGY G, EMODY L, PÁL T. Strategies for the development of vaccines conferring broad-spectrum protection[J]. International Journal of Medical Microbiology: IJMM, 2008, 298(5/6): 379-395.
97	SÁNCHEZ-SAMPEDRO L, PERDIGUERO B, MEJÍAS-PÉREZ E, et al. The evolution of poxvirus vaccines[J]. Viruses, 2015, 7(4): 1726-1803.
98	CUNNINGHAM A L, GARÇON N, LEO O, et al. Vaccine development: from concept to early clinical testing[J]. Vaccine, 2016, 34(52): 6655-6664.
99	LI Z H, SONG S, HE M Z, et al. Rational design of a triple-type human papillomavirus vaccine by compromising viral-type specificity[J]. Nature Communications, 2018, 9: 5360.
100	DE GROOT A S, MOISE L, TERRY F, et al. Better epitope discovery, precision immune engineering, and accelerated vaccine design using immunoinformatics tools[J]. Frontiers in Immunology, 2020, 11: 442.
101	ANTIA R, AHMED H, BULL J J. Directed attenuation to enhance vaccine immunity[J]. PLoS Computational Biology, 2021, 17(2): e1008602.
102	HAMMARLUND E, LEWIS M W, HANSEN S G, et al. Duration of antiviral immunity after smallpox vaccination[J]. Nature Medicine, 2003, 9(9): 1131-1137.

[1]	Zhidian DIAO, Xixian WANG, Qing SUN, Jian XU, Bo MA. Advances and applications of single-cell Raman spectroscopy testing and sorting equipment [J]. Synthetic Biology Journal, 2023, 4(5): 1020-1035.
[2]	Hui LU, Fangli ZHANG, Lei HUANG. Establishment of iBioFoundry for synthetic biology applications [J]. Synthetic Biology Journal, 2023, 4(5): 877-891.
[3]	Zhonghu BAI, He REN, Jianqi NIE, Yang SUN. The recent progresses and applications of in-parallel fermentation technology [J]. Synthetic Biology Journal, 2023, 4(5): 904-915.
[4]	Yujie WU, Xinxin LIU, Jianhui LIU, Kaiguang Yang, Zhigang SUI, Lihua ZHANG, Yukui ZHANG. Research progress of strain screening and quantitative analysis of key molecules based on high-throughput liquid chromatography and mass spectrometry [J]. Synthetic Biology Journal, 2023, 4(5): 1000-1019.
[5]	Zhehui HU, Juan XU, Guangkai BIAN. Application of automated high-throughput technology in natural product biosynthesis [J]. Synthetic Biology Journal, 2023, 4(5): 932-946.
[6]	Huan LIU, Qiu CUI. Advances and applications of ambient ionization mass spectrometry in screening of microbial strains [J]. Synthetic Biology Journal, 2023, 4(5): 980-999.
[7]	Yannan WANG, Yuhui SUN. Base editing technology and its application in microbial synthetic biology [J]. Synthetic Biology Journal, 2023, 4(4): 720-737.
[8]	Wanqiu LIU, Xiangyang JI, Huiling XU, Yicong LU, Jian LI. Cell-free protein synthesis system enables rapid and efficient biosynthesis of restriction endonucleases [J]. Synthetic Biology Journal, 2023, 4(4): 840-851.
[9]	Meili SUN, Kaifeng WANG, Ran LU, Xiaojun JI. Rewiring and application of Yarrowia lipolytica chassis cell [J]. Synthetic Biology Journal, 2023, 4(4): 779-807.
[10]	Zhi SUN, Ning YANG, Chunbo LOU, Chao TANG, Xiaojing YANG. Rational design for functional topology and its applications in synthetic biology [J]. Synthetic Biology Journal, 2023, 4(3): 444-463.
[11]	Qilong LAI, Shuai YAO, Yuguo ZHA, Hong BAI, Kang NING. Microbiome-based biosynthetic gene cluster data mining techniques and application potentials [J]. Synthetic Biology Journal, 2023, 4(3): 611-627.
[12]	Qiaozhen MENG, Fei GUO. Applications of foldability in intelligent enzyme engineering and design: take AlphaFold2 for example [J]. Synthetic Biology Journal, 2023, 4(3): 571-589.
[13]	Sheng WANG, Zechen WANG, Weihua CHEN, Ke CHEN, Xiangda PENG, Fafen OU, Liangzhen ZHENG, Jinyuan SUN, Tao SHEN, Guoping ZHAO. Design of synthetic biology components based on artificial intelligence and computational biology [J]. Synthetic Biology Journal, 2023, 4(3): 422-443.
[14]	Hailong LV, Jian WANG, Hao LV, Jin WANG, Yong XU, Dayong GU. Synthetic biology for next-generation genetic diagnostics [J]. Synthetic Biology Journal, 2023, 4(2): 318-332.
[15]	Mengdan MA, Yuchen LIU. Potential application of synthetic biology in disease information recording and real-time monitoring [J]. Synthetic Biology Journal, 2023, 4(2): 301-317.