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Cited by 745

The DOI matches its Crossref record; 30 of the 30 references it cites were found in a registry or an index.

Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector

Open access
Liu, Ziqing, Chen, Olivia, Wall, J. Blake Joseph, Zheng, Michael, Zhou, Yang, Wang, Li, Ruth Vaseghi, Haley, Qian, Li, Liu, Jiandong·2017·Scientific Reports

Abstract

Cloning of multiple genes in a single vector has greatly facilitated both basic and translational studies that require co-expression of multiple factors or multi-units of complex protein. Many strategies have been adopted, among which 2A “self-cleaving” peptides have garnered increased interest for their polycistronic nature, small size and high “cleavage” efficiency. However, broad application of 2 A peptides is limited by the lack of systematic comparison of different 2As alone or in combination. Here we characterized the effect of varying gene position and 2As on the expression of proteins encoded in bi-, tri-, or quad-cistronic constructs. Using direct cardiac reprogramming as an example, we further determined the effect of varied 2As on the efficiency of fluorescent cell labeling and cell fate conversion. We found that the expression of fluorophores decreased as it was moved towards the end of the construct while reprogramming was most efficient with the fluorophore at the second position. Moreover, quad-cistronic TPE2A constructs resulted in more efficient reprogramming than 3P2A or PTE2A constructs. We expect that the bi-, tri-, and quad-cistronic vectors constructed here and our results on protein expression ratios from different 2A constructs could serve to guide future utilization of 2A peptides in basic research and clinical applications.

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Metadata

Authors
Liu, Z., Chen, O., Wall, J. B. J., Zheng, M., Zhou, Y., Wang, L., Ruth Vaseghi, H., Qian, L., & Liu, J.
Venue
Scientific Reports
Published
2017-05-19
Volume
7
Issue
1
Pages
2193-2193
ISSN
2045-2322 (ISSN-L)
OpenAlex
W2615853094
Cited by
745
References
30
FWCI
12.71
Type
journal article
Language
en
Open access
gold

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CoChat. (2026). Citation check: Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. https://app.cochat.ai/papers/10.1038/s41598-017-02460-2
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Liu, Z., Chen, O., Wall, J. B. J., Zheng, M., Zhou, Y., Wang, L., Ruth Vaseghi, H., Qian, L., & Liu, J. (2017). Systematic comparison of 2A peptides for cloning multi-genes in a polycistronic vector. Scientific Reports. https://doi.org/10.1038/s41598-017-02460-2

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