dc.contributor.author | He, Jiening | |
dc.contributor.author | Huang, Lian | |
dc.contributor.author | Qiu, Huiling | |
dc.contributor.author | Li, Jiexuan | |
dc.contributor.author | Luo, Lan | |
dc.contributor.author | Li, Yanjiao | |
dc.contributor.author | Tian, Shengli | |
dc.contributor.author | Kang, Kang | |
dc.contributor.author | Luo, Jun | |
dc.contributor.author | Liu, Lin | |
dc.contributor.author | Gou, Deming | |
dc.date.accessioned | 2019-08-21T22:01:21Z | |
dc.date.available | 2019-08-21T22:01:21Z | |
dc.date.issued | 2015-12-08 | |
dc.identifier | oksd_he_anewdesignofale_2015 | |
dc.identifier.citation | He, J., Huang, L., Qiu, H., Li, J., Luo, L., Li, Y., ... Gou, D. (2015). A new design of a lentiviral shRNA vector with inducible co-expression of ARGONAUTE 2 for enhancing gene silencing efficiency. Cell and Bioscience, 5, Article 67. https://doi.org/10.1186/s13578-015-0058-2 | |
dc.identifier.uri | https://hdl.handle.net/11244/321182 | |
dc.description.abstract | Background: RNA interference (RNAi) is a robust tool for inhibiting specific gene expression, but it is limited by the uncertain efficiency of siRNA or shRNA constructs. It has been shown that the overexpression of ARGONAUTE 2 (AGO2) protein increases silencing efficiency. However, the key elements required for AGO2-mediated enhancement of gene silencing in lentiviral vector has not been well studied. | |
dc.description.abstract | Results: To explore the application of AGO2-based shRNA system in mammalian cells, we designed shRNA vectors targeting the EGFP reporter gene and evaluated the effects of various factors on silencing efficiency including stem length, loop sequence, antisense location as well as the ratio between AGO2 and shRNA. We found that 19 ~ 21-bp stem and 6- or 9-nt loop structure in the sense-loop-antisense (S-L-AS) orientation was an optimal design in the AGO2-shRNA system. Then, we constructed a single lentiviral vector co-expressing shRNA and AGO2 and demonstrated that the simultaneous expression of shRNA and AGO2 can achieve robust silencing of exogenous DsRed2 and endogenous ID1 and P65 genes. However, the titers of packaged lentivirus from constitutive expression of AGO2 vector were extremely low, severely limiting its broad application. For the first time, we demonstrated that the problem can be significantly improved by using the inducible expression of AGO2 lentiviral system. | |
dc.description.abstract | Conclusions: We reported a novel lentiviral vector with an optimal design of shRNA and inducible AGO2 overexpression which provides a new tool for RNAi research. | |
dc.format | application/pdf | |
dc.language | en_US | |
dc.publisher | BioMed Central | |
dc.rights | This material has been previously published. In the Oklahoma State University Library's institutional repository this version is made available through the open access principles and the terms of agreement/consent between the author(s) and the publisher. The permission policy on the use, reproduction or distribution of the material falls under fair use for educational, scholarship, and research purposes. Contact Digital Resources and Discovery Services at lib-dls@okstate.edu or 405-744-9161 for further information. | |
dc.title | New design of a lentiviral shRNA vector with inducible co-expression of ARGONAUTE 2 for enhancing gene silencing efficiency | |
osu.filename | oksd_he_anewdesignofale_2015.pdf | |
dc.description.peerreview | Peer reviewed | |
dc.identifier.doi | 10.1186/s13578-015-0058-2 | |
dc.description.department | Physiological Sciences | |
dc.type.genre | Article | |
dc.type.material | Text | |
dc.subject.keywords | ago2 | |
dc.subject.keywords | lentivirus | |
dc.subject.keywords | rnai | |
dc.subject.keywords | shrna | |