IDT MiniGene constructs were used as microRNA (miR) sponges against miR-17, miR-18a, miR-19, and miR-92. The sequences were also designed with 5′ Xho1 and 3′ EcoR1 sites for subcloning into other vectors by restriction digest.
IDT MiniGene synthetic genes were engineered for use in transcription factor binding assays. Mutant sites were created for Sp1, HNF-1, and NF-Y and the MiniGene synthetic genes were provided in pIDTSMART-KAN vectors.
A portion of the second exon of herpes simplex virus, immediate early protein ICP0, was synthesized as a MiniGene synthetic gene. The MiniGene synthetic gene construct was subcloned into a secondary vector system and used to generate RNA for use as a standard in RT-qPCR.
IDT MiniGene constructs were designed as IRF2BP2 (Interferon regulatory factor 2 binding protein 2) mutants: 355 GAC GAC GAC GAC 358 (aspartic acids), TCT 360 TAT (S360A), and TCT 360 GAT (S360D). The MiniGene cosnstructs were flanked by endogenous PstI and NcoI in IRF2BP2 for subcloning by restriction digest.
An IDT MiniGene expressing wild-type survival motor neuron from SMN2, and a second SMN2 MiniGene with a silent mutation in the siRNA recognition sequence, were used to study the effects of siRNA knockdown, and rescue, of SMN1/2 on gene splicing. IDT DsiRNA Duplexes were also used for the siRNA knockdown experiments.