Pol I-dependent shRNA induces species-specific gene silencing

Copyright information:

Taken from "Development of a species-specific RNA polymerase I-based shRNA expression vector"

Nucleic Acids Research 2006;35(2):e10-e10.

Published online 07 Dec 2006

PMCID:PMC1802596.

© 2006 The Author(s).

() Twenty four-well plates of HT22 mouse hippocampal cells were transfected with 80 ng/well of plasmid that directs the expression of firefly (pGL3-Con, Promega), 8 ng/well of plasmid expressing (pRL-SV40, Promega) luciferase and 800 ng/well of the indicated DNA. Luciferase activities were assayed 48 h after transfection using the dual luciferase assay. Ratios of firefly to luciferase activities were normalized to a control transfected with vector pcDNA3, and the control ratio was set to 1. The average of two independent experiments is shown, and each experiment was performed in duplicate; error bars indicate SD. pE3SP-MCS and pU6 are empty vectors for corresponding shLuc-expressing plasmids pE3SP-shLuc and pU6-shLuc, respectively. < 0.001 Statistical significance was evaluated using the Student's test. () Species specificity of Pol I-dependent gene silencing. Twenty four-well plates of HT22 mouse hippocampal cells or HEK293 human embryonic kidney cells were transfected and the luciferase expression was measured as in (A). Ratios of firefly to luciferase activities were normalized to a control transfected with pBluescript vector. The average of two independent experiments is shown, and each experiment was performed in duplicate; error bars indicate SD. < 0.001, < 0.005. () Species specificity of Pol I-dependent gene silencing. Twenty four-well plates of HeLa human epithelial cells, Neuro-2a mouse neuroblastoma cells or HN9 mouse embryonic hippocampal cells were transfected, and luciferase expression was measured as in (A). Ratios of firefly to luciferase activities are expressed as a percentage of the control sample that was transfected with the accordant empty expression vector (pU6 for pU6-shLuc and pE3SP-MCS for pE3SP-shLuc). The average of two independent experiments is shown, and each experiment was performed in triplicate; error bars indicate SDs. Statistical analysis was done using the Student's test and asterisks indicate significant difference from the empty vector control. < 0.001.