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An Argonaute phosphorylation cycle promotes microRNA-mediated silencing.
Golden RJ, Chen B, Li T, Braun J, Manjunath H, Chen X, Wu J, Schmid V, Chang TC, Kopp F, Ramirez-Martinez A, Tagliabracci VS, Chen ZJ, Xie Y, Mendell JT
Nature. 2017 Feb 9;542(7640):197-202. doi: 10.1038/nature21025. Epub 2017 Jan 23.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
91975MSCV-EGFPEGFP reporter cell line control (no miR-19 sites)
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91976MSCV-EGFP-Bulged-miR-19EGFP reporter cell line with eight bulged miR-19 sites
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91977LcV2-HygroLentiCRISPR v2 with Hygro cloned in place of Puro
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91978pLJM1-FH-AGO2-WTExpresses FLAG-HA-AGO2 (WT)
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91979pLJM1-FH-AGO2-5xAExpresses FLAG-HA-AGO2 [S824A, S828A, T830A, S831A, S834A (5xA)]
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91980pLJM1-EmptyEmpty backbone
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91981pLJM1-FH-AGO2-5xEExpresses FLAG-HA-AGO2 [S824E, S828E, T830E, S831E, S834E (5xE)]
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91982pLJM1-FH-AGO2-S828AExpresses FLAG-HA-AGO2 (S828A)
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91983pLJM1-FH-AGO2-D358KExpresses FLAG-HA-AGO2 (D358K)
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91984pLJM1-FH-AGO2-R438EExpresses FLAG-HA-AGO2 (R438E)
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91985pLJM1-FH-AGO2-K525EExpresses FLAG-HA-AGO2 (K525E)
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91986MSCV-Puro-FH-AGO2-WTExpresses FLAG-HA-AGO2 (WT)
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91987MSCV-Puro-FH-AGO2-S824AExpresses FLAG-HA-AGO2 (S824A)
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91988MSCV-Puro-FH-AGO2-T830AExpresses FLAG-HA-AGO2 (T830A)
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91989MSCV-Puro-FH-AGO2-S831AExpresses FLAG-HA-AGO2 (S831A)
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91990MSCV-Puro-FH-AGO2-S834AExpresses FLAG-HA-AGO2 (S834A)
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91991MSCV-Puro-FH-AGO2-S828AExpresses FLAG-HA-AGO2 (S828A)
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91992MSCV-Puro-FH-AGO2-5xAExpresses FLAG-HA-AGO2 [S824A, S828A, T830A, S831A, S834A (5xA)]
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91993MSCV-Puro-FH-AGO2-5xEExpresses FLAG-HA-AGO2 [S824E, S828E, T830E, S831E, S834E (5xE)]
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91994MSCV-Puro-FH-AGO2-S824EExpresses FLAG-HA-AGO2 (S824E)
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91995MSCV-Puro-FH-AGO2-S828EExpresses FLAG-HA-AGO2 (S828E)
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91996MSCV-Puro-FH-AGO2-T830EExpresses FLAG-HA-AGO2 (T830E)
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91997MSCV-Puro-FH-AGO2-S831EExpresses FLAG-HA-AGO2 (S831E)
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91998MSCV-Puro-FH-AGO2-S834EExpresses FLAG-HA-AGO2 (S834E)
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91999MSCV-Puro-FH-AGO2-S824E-S828EExpresses FLAG-HA-AGO2 (S824E-S828E)
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92000MSCV-Puro-FH-AGO2-S824E-S831EExpresses FLAG-HA-AGO2 (S824E-S831E)
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92001MSCV-Puro-FH-AGO2-S828E-S831EExpresses FLAG-HA-AGO2 (S828E-S831E)
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92002MSCV-Puro-FH-AGO2-S824E-S828E-S831EExpresses FLAG-HA-AGO2 (S824E-S828E-S831E)
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92003MSCV-Puro-FH-AGO2-S824A-T830AExpresses FLAG-HA-AGO2 (S824A-T830A)
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92004pGL3-BoxBExpresses firefly luciferase with BoxB sequences in 3' UTR
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92005pcDNA3.1+_Lambda-N-HA-PeptideExpresses Lambda-N-HA peptide
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92006pcDNA3.1+_FH-AGO2-WTExpresses FLAG-HA-AGO2 (WT)
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92007pcDNA3.1+_FH-AGO2-5xAExpresses FLAG-HA-AGO2 [S824A, S828A, T830A, S831A, S834A (5xA)]
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92008pcDNA3.1+_FH-AGO2-5xEExpresses FLAG-HA-AGO2 [S824E, S828E, T830E, S831E, S834E (5xE)]
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92009pcDNA3.1+_LN-HA-AGO2-WTExpresses Lambda-N-HA-AGO2 (WT)
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92010pcDNA3.1+_LN-HA-AGO2-5xAExpresses Lambda-N-HA-AGO2 [S824A, S828A, T830A, S831A, S834A (5xA)]
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92011pcDNA3.1+_LN-HA-AGO2-5xEExpresses Lambda-N-HA-AGO2 [S824E, S828E, T830E, S831E, S834E (5xE)]
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92012pcDNA3.1+_V5-ANKRD52Expresses V5-ANKRD52
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92013pCAGIG_V5-PPP6CExpresses V5-PPP6C, includes IRES-Emerald
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92014pCAGImC_V5-CSNK1A1Expresses V5-CSNK1A1, includes IRES-mCherry
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92015pCAGImC_EmptyEmpty backbone, includes IRES-mCherry
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