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Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display.
Shechner DM, Hacisuleyman E, Younger ST, Rinn JL
Nat Methods. 2015 Jul;12(7):664-70. doi: 10.1038/nmeth.3433. Epub 2015 Jun 1. PubMed Article

Plasmids from Article

ID Plasmid Purpose
68414pLenti_9xD3B_Cluc_NormCRISPR-Display Cypridina Luciferase-2A-Venus YFP "normalizer," lentiviral format.
68415pTrans_9xD3B_Cluc_NormCRISPR-Display Cypridina Luciferase-2A-Venus YFP "normalizer," transient format.
68416pEF_dCas9Transient expression of Sp-dCas9 in mammalian cells, under an EF1-alpha promoter.
68417pEF_dCas9-VP64Transient expression of Sp-dCas9 fused to the VP64 transcription activator, in mammalian cells, under an EF1-alpha promoter.
68418pUBC_HA_2xPP7-VP64Transient expression of two tandem copies of the non-aggregating delta-FG mutant, fused to the VP64 transcription activator, in mammalian cells, under a UBC promoter
68419pCMV_mCherry_2A_HA_2xMS2-VP64Transient expression of humanized MS2 (V75E/A81G), fused to the VP64 transcrption activator, in mammalian cells, under a CMV promoter
68420pUBC_V5_MS2-mCherryTransient expression of humanized MS2 (V75E/A81G), fused to mCherry in mammalian cells, under a UBC promoter
68421pUBC_HA_L7Ae-VP64Transient expression of humanized A. fulgidus L7Ae, fused to the VP64 transcription activator, in mammalian cells, under a UBC promoter
68422pU6_(GLuc)_sgRNATransient expression of a minimal sgRNA targeting the GLuc reporter, in mammalian cells. U6 promoter
68423pU6_(GLuc)_TOP1Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. U6 promoter
68424pU6_(GLuc)_INT[3xPP7_SL]Transient expression of an "INT" construct_bearing three PP7 Stem-loops, targeting the GLuc reporter, in mammalian cells. U6 promoter
68425pU6_(GLuc)_INT[S1Apt]Transient expression in mammalian cells of an "INT" construct_bearing the "S1" streptavidin aptamer, targeting the GLuc reporter. U6 promoter
68426pU6_(GLuc)_INT[3xMS2_SL]Transient expression in mammalian cells of an "INT" construct_bearing three MS2 Stem-loops, targeting the GLuc reporter. U6 promoter
68428pU6_(GLuc)_INT[GFPApt]Transient expression in mammalian cells of an "INT" construct_bearing the GFP aptamer, targeting the GLuc reporter. U6 promoter
68429pU6_(GLuc)_INT[3xK-T]Transient expression in mammalian cells of an "INT" construct_bearing three kink-turns, targeting the GLuc reporter. U6 promoter
68430pU6_(GLuc)_INT[Spinach2]Transient expression in mammalian cells of an "INT" construct_bearing the "Spinach2" aptamer, targeting the GLuc reporter. U6 promoter
68431pU6_(GLuc)_INT[BoBS]Transient expression in mammalian cells of an "INT" construct_bearing the "Bunch of Baby Spinach" (or "BoBS") construct, targeting the GLuc reporter. U6 promoter
68432pU6_(GLuc)_INT[P4-P6(3xPP7)]Transient expression in mammalian cells of an "INT" construct_bearing P4_P6 (internally appended with three PP7 stem-loops), targeting the GLuc reporter. U6 promoter
68433pU6_(Gluc)_INT(GenPurpClon)General Purpose cloning vector for "INT"-like constructs under U6-promoter expression.
68434pCMV/3' Box_(GLuc)_TOP1Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. CMV/3' Box expression backbone.
68435pCMV/3' Box_(GLuc)_TOP2Transient expression of the "TOP2" construct, targeting the GLuc reporter, in mammalian cells. CMV/3' Box expression backbone.
68436pCMV/3' Box_(GLuc)_INTTransient expression of an "INT" construct_bearing three PP7 Stem-loops, targeting the GLuc reporter, in mammalian cells. CMV/3' Box expression backbone.
68437pU1/Sm/3' Box_(GLuc)_TOP1Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. U1Pro/SmBox/3' Box expression backbone.
68438pU1/Sm/3' Box_(GLuc)_INTTransient expression of an "INT" construct_bearing three PP7 Stem-loops, targeting the GLuc reporter, in mammalian cells. U1Pro/SmBox/3' Box expression backbone.
68439pCMV/MASC_(GLuc)_TOP1Transient expression of the "TOP1" construct, targeting the GLuc reporter, in mammalian cells. CMV/MASC expression backbone.
68440pCMV/MASC_(GLuc)_INTTransient expression of an "INT" construct_bearing three PP7 Stem-loops, targeting the GLuc reporter, in mammalian cells. CMV/ MASC expression backbone.

Antibodies from Article