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URL: http://www.addgene.org/67879
Proper Citation: RRID:Addgene_67879
Insert Name: actin prom-dsRed-scramble shRNA
Organism: Gallus gallus
Bacterial Resistance: Chloramphenicol and Ampicillin
Defining Citation: PMID:25706476
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pUC; Vector Types:Mammalian Expression, AAV, RNAi; Bacterial Resistance:Chloramphenicol and Ampicillin
Comments: To express shRNAs, we used the pPRIME system. This generates micro-RNA-30 (mir30)-derived shRNAs and co-expression of a marker protein, such as dsRED, from the same transcript. The shRNA hairpin sequences were placed into the mir30 site of pPRIME-cmv-dsRed-FF3 and then cloned into the AAV vector (Zhang et al 2015 Nat Neurosci), SbfI(or PstI) + PacI will retrieve the 2.1kb insert (dsRed-scramble shRNA-chloramphenicol) and the 5.1kb AAV-CBA backbone, which is ampicillin resistant. The SbfI site in the middle of dsRed has been silenced. The construct can be linearised with XhoI.
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Source: Addgene
