ArticleInternational journal of biochemistry and molecular biology2013
Insert sequence length determines transfection efficiency and gene expression levels in bicistronic mammalian expression vectors.
Article in International journal of biochemistry and molecular biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
What it found
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Who cites it
6 citing papers in PubMed.
- Review
- Guidelines for minimal reporting requirements, design and interpretation of experiments involving the use of eukaryotic dual gene expression reporters (MINDR).Nature structural & molecular biology · 2025Review
- Development, Characterization, and Application of Two Reporter-Expressing Recombinant Zika Viruses.Viruses · 2020Article
- Stop codon readthrough contexts influence reporter expression differentially depending on the presence of an IRES.Wellcome open research · 2020Article
- The use of a viral 2A sequence for the simultaneous over-expression of both the vgf gene and enhanced green fluorescent protein (eGFP) in vitro and in vivo.Journal of neuroscience methods · 2015Article
- Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Bicistronic expression vectors have been widely used for co-expression studies since the initial discovery of the internal ribosome entry site (IRES) about 25 years ago. IRES sequences allow the 5' cap-independent initiation of translation of multiple genes on a single messenger RNA strand. Using a commercially available mammalian expression vector containing an IRES sequence with a 3' green fluorescent protein fluorescent marker, we found that sequence length of the gene of interest expressed 5' of the IRES site influences both expression of the 3' fluorescent marker and overall transfection efficiency of the vector construct. Furthermore, we generated a novel construct expressing two distinct fluorescent markers and found that high expression of one gene can lower expression of the other. Observations from this study indicate that caution is warranted in the design of experiments utilizing an IRES system with a short 5' gene of interest sequence (<300 bp), selection of single cells based on the expression profile of the 3' optogenetic fluorescent marker, and assumptions made during data analysis.
Indexed as
Identifiers
24380024PMC3867706What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.