Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform

Thao, Tran Thi Nhu; Labroussaa, Fabien; Ebert, Nadine; V’kovski, Philip; Stalder, Hanspeter; Portmann, Jasmine; Kelly, Jenna N.; Steiner, Silvio; Holwerda, Melle; Kratzel, Annika; Gultom, Mitra; Schmied, Kimberly; Laloli, Laura; Hüsser, Linda; Wider, Manon; Pfaender, Stephanie; Hirt, Dagny; Cippà, Valentina; Crespo-Pomar, Silvia; Schröder, Simon; Muth, Doreen; Niemeyer, Daniela; Corman, Victor M.; Müller, Marcel A.; Drosten, Christian; Dijkman, Ronald; Jores, Joerg; Thiel, Volker

doi:10.1038/s41586-020-2294-9

Cited by 388 publications

(251 citation statements)

References 35 publications

Supporting

Mentioning

239

Contrasting

Unclassified

Order By: Relevance

“…To assess the impact of S-614G in the context of virus infection we generated an isogenic D614G virus pair based on our reverse genetics system for SARS-CoV-2 10 . The molecular clone is based on the Wuhan-Hu-1 isolate possessing the S-614D variant (SARS-CoV-2 S-614D ) 10,11 . The sequence of the isogenic S-614G variant was engineered to have an A to G nucleotide change at position 23,403 to encode a glycine at the S protein position 614.…”

Section: Resultsmentioning

confidence: 99%

“…To introduce the 614G mutation to the Spike gene, PCR mutagenesis (Supplementary Table 1) was performed on the pUC57 plasmid containing SARS-CoV-2 fragment 10 10 using Q5® Site-Directed Mutagenesis Kit (New England BioLab). Both D614 and G614 infectious cDNA clones were generated using in-yeast TAR cloning method as describe previously 10 . In vitro transcription was performed for EagI-cleaved YACs and PCR-amplified SARS-CoV-2 N gene using the T7 RiboMAX Large Scale RNA production system (Promega) as described previously 26 .…”

Section: Methodsmentioning

confidence: 99%

“…Characterization of virus growth kinetics in Vero E6 was performed as described previously 10 . Twenty-four hours before infection, cells were seeded in a 24-well plate at a density of 2.0 × 10 5 cells per ml.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

SARS-CoV-2 spike D614G variant confers enhanced replication and transmissibility

Zhou

Thao

Hoffmann

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

During the evolution of SARS-CoV-2 in humans a D614G substitution in the spike (S) protein emerged and became the predominant circulating variant (S-614G) of the COVID-19 pandemic. However, whether the increasing prevalence of the S-614G variant represents a fitness advantage that improves replication and/or transmission in humans or is merely due to founder effects remains elusive. Here, we generated isogenic SARS-CoV-2 variants and demonstrate that the S-614G variant has (i) enhanced binding to human ACE2, (ii) increased replication in primary human bronchial and nasal airway epithelial cultures as well as in a novel human ACE2 knock-in mouse model, and (iii) markedly increased replication and transmissibility in hamster and ferret models of SARS-CoV-2 infection. Collectively, our data show that while the S-614G substitution results in subtle increases in binding and replication in vitro, it provides a real competitive advantage in vivo, particularly during the transmission bottle neck, providing an explanation for the global predominance of S-614G variant among the SARS-CoV-2 viruses currently circulating.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

SARS-CoV-2 spike D614G variant confers enhanced replication and transmissibility

Zhou

Thao

Hoffmann

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…On 21 February, a Swiss lab announced that it had synthesized the SARS-CoV-2 genome from synthetic DNA constructs ordered on 14 January using publicly available GSD. In their preprint published on the bioRxiv server, the research team detailed the synthesis of multiple RNA viruses, including SARS-CoV-2, using their yeastbased reverse genetics platform (11). The team reported difficulties in creating two of the 14 synthetic DNA fragments to synthesize SARS-CoV-2, requiring a physical virus sample isolated from a COVID-19 patient in Germany in order to recover infectious virus particles (11).…”

Section: Samples Without Samples: Dematerialsization and Rematerialsizamentioning

confidence: 99%

“…In their preprint published on the bioRxiv server, the research team detailed the synthesis of multiple RNA viruses, including SARS-CoV-2, using their yeastbased reverse genetics platform (11). The team reported difficulties in creating two of the 14 synthetic DNA fragments to synthesize SARS-CoV-2, requiring a physical virus sample isolated from a COVID-19 patient in Germany in order to recover infectious virus particles (11). Even though the first SARS-CoV-2 genetic sequence was shared early, it is clear that these developments in synthetic biology could make governments reluctant to share GSD on openly accessible databases if it means they could miss out on benefits that might otherwise be gained by enforcing their domestic ABS laws.…”

Section: Samples Without Samples: Dematerialsization and Rematerialsizamentioning

confidence: 99%

Policy opportunities to enhance sharing for pandemic research

Rourke

Eccleston-Turner

Phelan

et al. 2020

Science

View full text Add to dashboard Cite

show abstract

DNA‐Based Materials Inspired by Natural Extracellular DNA

Qin

Wang

Zhang

et al. 2023

Adv Funct Materials

View full text Add to dashboard Cite

The advent of biotechnology has expedited the understanding of the biochemistry of deoxyribonucleic acids (DNA). In the past, DNA are thought to be present only in cell nucleus as bearers of the genetic code. With the identification of extracellular DNA in circulating body fluids, DNA are now utilized, at least experimentally, for diagnosis and treatment of diseases. Extracellular DNA of host origin trigger immune responses, and are closely linked to autoimmune disease, cancer-related inflammation, bacteria adhesion and thrombosis. Recent advancements in DNA nanotechnology have led to the development of a series of DNA-based materials for treating diseases because of their structural programmability. Current discussions on biosafety and immunogenicity of artificial DNA materials are insufficient. This issue severely restricts the clinical translation of these novel biotechnologies. The present review attempts to bridge the gap between natural extracellular DNA and their derivatives, DNA-based materials. The pathological attributes of endogenous extracellular DNA motivate the design of targeting DNA materials. In addition, the fate of exogenous DNA in the host inspires the optimization of DNA materials in reducing immune rejection. These bioinspired strategies provide the blueprint for utilizing DNA materials in the management of diseases that are currently challenging to diagnose or treat.

show abstract

Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform

Cited by 388 publications

References 35 publications

SARS-CoV-2 spike D614G variant confers enhanced replication and transmissibility

SARS-CoV-2 spike D614G variant confers enhanced replication and transmissibility

Policy opportunities to enhance sharing for pandemic research

DNA‐Based Materials Inspired by Natural Extracellular DNA

Contact Info

Product

Resources

About