Development of Host‐Orthogonal Genetic Systems for Synthetic Biology

Zhang, Yang; Ding, Wentao; Wang, Zhihui; Zhao, Huimin; Shi, Shuobo

doi:10.1002/adbi.202000252

Cited by 8 publications

(5 citation statements)

References 163 publications

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“…Precise and reliable control over gene expression is one of the most fundamental requirements of synthetic biology (1). Consequently, there has been considerable effort towards identifying myriad genetic elements that enable researchers to regulate the strength and timing of transcription across all domains of life (2)(3)(4). The end result of these efforts are often consolidated families of plasmid vectors that facilitate advanced genetic engineering, such as the BglBrick family of plasmids for E. coli (5,6) and the jStack vectors used in multiple plant species (7).…”

Section: Introductionmentioning

confidence: 99%

The pGinger family of expression plasmids

Pearson

Thompson

et al. 2023

Preprint

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The pGinger suite of expression plasmids comprises 43 plasmids that will enable precise constitutive and inducible gene expression in a wide range of gram-negative bacterial species. Constitutive vectors are composed of 16 synthbuetic constitutive promoters upstream of RFP, with a broad host range BBR1 origin and a kanamycin resistance marker. The family also has seven inducible promoters (Jungle Express, NahR, XylS, RhaS, Lac, LacUV5, and TetR) controlling RFP expression on BBR1/kanamycin plasmid backbones. For four of these inducible promoters (Jungle Express, NahR, Lac, and TetR), we created variants that utilize the RK2 origin and spectinomycin or gentamicin selection. Relevant RFP expression and growth data have been collected in the model bacterium Escherichia coli as well as Pseudomonas putida. All pGinger vectors are available via the Joint BioEnergy Institute (JBEI) Public Registry.

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Section: Introductionmentioning

confidence: 99%

The pGinger family of expression plasmids

Pearson

Thompson

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Precise and reliable control over gene expression is one of the most fundamental requirements of synthetic and molecular biology ( 1 ). Consequently, there has been considerable effort toward identifying myriad genetic elements that enable researchers to regulate the strength and timing of transcription across all domains of life ( 2 – 4 ). The results of these efforts are often consolidated families of plasmid vectors that facilitate advanced genetic engineering, such as the BglBrick family of plasmids for Escherichia coli ( 5 , 6 ) and the jStack vectors used in multiple plant species ( 7 ).…”

Section: Introductionmentioning

confidence: 99%

The pGinger Family of Expression Plasmids

et al. 2023

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Metabolic engineering and synthetic biology are predicated on the precise control of gene expression. As synthetic biology expands beyond model organisms, more tools will be required that function robustly in a wide range of bacterial hosts. The pGinger family of plasmids constitutes 43 plasmids that will enable both constitutive and inducible gene expression in a wide range of nonmodel Proteobacteria .

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“…The orthogonal biosystem played a critical role in on-going synthetic biology, consisting of the host-independent elements or molecules, for example, DNA, RNA, protein, or small chemicals. [1] The RNA polymerase derived from bacteriophage T7 and its cognate promoter were typical orthogonal transcription system. In addition to the specific recognition between T7 RNAP and T7 promoter, T7 system also has a number of other valuable characterizations, such as single-subunit structure, high processivity, and high transcription capacity.…”

Section: Introductionmentioning

confidence: 99%

Construction and Application of Orthogonal T7 Expression System in Eukaryote: An Overview

Wang

Yan

et al. 2022

Advanced Biology

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to its high efficiency and yield for protein synthesis. [12] Among the commonlyused protein expression systems in E. coli (Table 1), T7 expession system prepared more than 90% of proteins in protein data bank (PDB) due to its high yield and translation efficiency for target protein. [13] The T7 system had been applied extensively for target protein and mRNA synthesis in prokaryote, eukaryote, and cell free system. [2] UP to now, it was also becoming an enabling tool for on-going synthetic biology, metabolic engineering, biomedicine, enzyme engineering, and so on. [14][15][16][17] Although T7 expression system had been developed successfully in various prokaryotes, [2] the construction of T7 expression system in eukaryote was still on-going, which attributed to the innate feature of transcription-translation system in eukaryote, for example, in prokaryote, the transcription was coupled with translation, while in eukaryote, mRNA was synthesized and modified in nucleus, then exporting to cytoplasm for protein synthesis. Thus far, there was no systematic and complete review about T7 expression system construction in eukaryote. In the present paper, the development of T7 expression system in eukaryote was reviewed, including its construction in animal (mammalian cells, trypanosomatid protozoa, Xenopus oocytes, zebrafish), plant, and microorganism and its application in vaccine preparation and gene therapy. We also discussed the innate challenges of T7 expression system construction in eukaryote and its recent development in vaccine production and gene therapy. Animal Mammalian CellsDue to the pivotal role in life science research, clinical research, gene therapy, and vaccine production, mammalian cells were first selected as the eukaryotic host to construct T7 expression system; so far, mammalian cells have experienced a deep and systematic study on the construction of T7 expression system. In 1986, a vaccinia/T7 hybrid virus expression system (VT7 expression system) was constructed in CV-1 monkey kidney cells, in which two expression vectors co-infected host cells, including a recombinant vaccinia virus containing vaccinia promoter-driven T7 RNAP and a plasmid containing T7 promoter-driven target gene (Figure 1A). [26] Vaccinia is a cytoplasmic DNA virus, that is, the genome DNA of vaccinia exists in host cytoplasm. Vaccinia genome encodes all replication andThe T7 system is an orthogonal transcription-system, which is characterized by simplicity, higher efficiency, and higher processivity, and it is used for protein or mRNA synthesis in various biological-systems. In comparison with prokaryotes, the construction of the T7 expression system is still ongoing in eukaryotes, but it shows greatly applicable prospects. In the present paper, development of T7 expression system construction in eukaryotes is reviewed, including its construction in animal (mammalian cells, trypanosomatid protozoa, Xenopus oocytes, zebrafish), plant, and microorganism and its application in vaccine production and gene therapy. In addition, the...

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Development of Host‐Orthogonal Genetic Systems for Synthetic Biology

Cited by 8 publications

References 163 publications

The pGinger family of expression plasmids

The pGinger family of expression plasmids

The pGinger Family of Expression Plasmids

Construction and Application of Orthogonal T7 Expression System in Eukaryote: An Overview

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