Noncoding RNA gene silencing through genomic integration of RNA destabilizing elements using zinc finger nucleases

Knockdown of the abundant ncRNA MALAT1. (A) The relative expression levels of GAPDH, RPLP0, ACTB, and MALAT1 in A549 cells were determined via qRT–PCR and analyzed using the 2^(-ddCt) method (Livak and Schmittgen 2001). RN7SL1 was used as reference gene. Shown is the mean of measurements from two experiments (×10⁻⁷) and the standard deviation (SD). (B) Schematic overview of the siRNA and qPCR primer position in the MALAT1 transcript. (C) Targeting of MALAT1 with seven different siRNAs yielded a knockdown to, at maximum, 13% remaining expression. The transcript level was determined via qRT–PCR and analyzed using the 2^(-ddCt) method (Livak and Schmittgen 2001) with RN7SL1 as the reference gene (mean + SD; n = 2).