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- W2000622419 endingPage "e76279" @default.
- W2000622419 startingPage "e76279" @default.
- W2000622419 abstract "The ability to express exogenous cDNAs while suppressing endogenous genes via RNAi represents an extremely powerful research tool with the most efficient non-transient approach being accomplished through stable viral vector integration. Unfortunately, since traditional restriction enzyme based methods for constructing such vectors are sequence dependent, their construction is often difficult and not amenable to mass production. Here we describe a non-sequence dependent Gateway recombination cloning system for the rapid production of novel lentiviral (pLEG) and retroviral (pREG) vectors. Using this system to recombine 3 or 4 modular plasmid components it is possible to generate viral vectors expressing cDNAs with or without inhibitory RNAs (shRNAmirs). In addition, we demonstrate a method to rapidly produce and triage novel shRNAmirs for use with this system. Once strong candidate shRNAmirs have been identified they may be linked together in tandem to knockdown expression of multiple targets simultaneously or to improve the knockdown of a single target. Here we demonstrate that these recombinant vectors are able to express cDNA and effectively knockdown protein expression using both cell culture and animal model systems." @default.
- W2000622419 created "2016-06-24" @default.
- W2000622419 creator A5014504951 @default.
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- W2000622419 creator A5047958594 @default.
- W2000622419 creator A5054355892 @default.
- W2000622419 creator A5085929194 @default.
- W2000622419 creator A5088100193 @default.
- W2000622419 date "2013-10-11" @default.
- W2000622419 modified "2023-10-18" @default.
- W2000622419 title "A Modular Lentiviral and Retroviral Construction System to Rapidly Generate Vectors for Gene Expression and Gene Knockdown In Vitro and In Vivo" @default.
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- W2000622419 doi "https://doi.org/10.1371/journal.pone.0076279" @default.
- W2000622419 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3795761" @default.
- W2000622419 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/24146852" @default.