In the 1970s, recombinant DNA technology enabled the engineering of artificial plasmids carrying foreign DNA of interest (2), which in the ensuing years propelled plasmids to become a central and ubiquitous reagent in the life sciences. Nowadays, plasmids are used mostly as gene delivery vectors in vitro and in vivo, either directly or as starting ...
For many decades, researchers have made their own customized plasmids in the lab to meet their specific research needs. Yet, not only can lab-made plasmids be constructed with mistakes, they are also subject to ongoing mutations while maintained in E. coli due to DNA replication errors (16), homologous recombination (17), and insertions of jumping ...
We found that approximately 15% of plasmids had significant design errors, and about 35% contained sequence errors in functional regions (excluding AAV ITRs) (Figure 1)
The high error rate of lab-made plasmids suggests that many labs lack the sophisticated and nuanced expertise needed to properly design vectors and furthermore, there is insufficient quality control of the plasmids being constructed and propagated in labs.
We therefore suggest that researchers should stay cautious of the functionality and sequence fidelity of plasmids obtained from public repositories.
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