The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.
Quantitative assessment on the cloning efficiencies of lentiviral transfer vectors with a unique clone site
Open accessAbstract
Lentiviral vectors (LVs) are powerful tools for transgene expression in vivo and in vitro. However, the construction of LVs is of low efficiency, due to the large sizes and lack of proper clone sites. Therefore, it is critical to develop efficient strategies for cloning LVs. Here, we reported a combinatorial strategy to efficiently construct LVs using EGFP, hPlk2 wild type (WT) and mutant genes as inserts. Firstly, site-directed mutagenesis (SDM) was performed to create BamH I site for the inserts; secondly, pWPI LV was dephosphorylated after BamH I digestion; finally, the amounts and ratios of the insert and vector DNA were optimized to increase monomeric ligation. Our results showed that the total percentage of positive clones was approximately 48%±7.6%. Using this method, almost all the vectors could be constructed through two or three minipreps. Therefore, our study provided an efficient method for constructing large-size vectors.
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Cites
In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector
Naldini, L., Blömer, U., Gallay, P., Ory, D., Mulligan, R., Gage, F. H., Verma, I. M., & Trono, D. (1996) · Science · doi:10.1126/science.272.5259.263 · Cited by 4,903
Induced Pluripotent Stem Cell Generation Using a Single Lentiviral Stem Cell Cassette
Sommer, C. A., Stadtfeld, M., Murphy, G. J., Hochedlinger, K., Kotton, D. N., & Mostoslavsky, G. (2009) · Stem Cells · doi:10.1634/stemcells.2008-1075 · Cited by 676
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Minnaert, A.-K., Vanluchene, H., Verbeke, R., Lentacker, I., De Smedt, S. C., Raemdonck, K., Sanders, N. N., & Remaut, K. (2021) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2021.113900 · Cited by 157
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Metadata
- Authors
- Zhang, G., & Tandon, A.
- Venue
- Scientific Reports
- Published
- 2012-05-22
- Volume
- 2
- Issue
- 1
- Pages
- 1-8
- ISSN
- 2045-2322 (ISSN-L)
- OpenAlex
- W1968368569
- Cited by
- 1,012
- References
- 2
- FWCI
- 61.29
- Type
- journal article
- Language
- en
- Open access
- gold
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