Journal: bioRxiv

Article Title: Dual-acting gene therapy targeting HIF1A and HIF2A by RNA interference mitigates retinal degeneration in two mouse models of AMD

doi: 10.1101/2024.11.29.626080

Figure Lengend Snippet: (A) Graphical representation of tissue changes leading to hypoxia and degenerative processes during aging and AMD. (B) Gene expression analysis of 661W cells stably expressing shCtrl, shHif1 or shHif2 after normoxic or hypoxic (24 h, 0.2% O 2 ) exposure. Values were normalized to beta Actin ( Actb ) and expressed relatively to normoxic 661W-shCtrl (set to 1). Shown are individual values (n=3) and means ± SD. Significance was tested using a one-way ANOVA with Tukey’s multiple comparison test. *: statistically significant; p-values of all comparisons are given in Table S1. (C) Western blot analysis for HIF1A, HIF2A and ACTB in 661W cells stably expressing shCtrl, shHif1 or shHif2 after normoxic and hypoxic exposure. (D) Schematic representation of the DNA constructs packaged into AAV vectors. AAV-shHif: the construct consists of the GRK1 promotor driving photoreceptor-specific expression of shHif1a embedded in the miR-E scaffold in the 3’UTR of GFP, followed by the VMD2 promotor controlling RPE-specific expression of shHif2a embedded in the miR-E scaffold in the 3’UTR of mCherry. AAV-shCtrl: a scrambled non-targeting control shRNA sequence replaced shHif1a and shHif2a. AAV-shHif1: the GRK1 promotor drives photoreceptor-specific expression of shHif1a embedded in the miR-E scaffold in the 3’UTR of GFP. AAV-shHif2: the VMD2 promotor controls RPE-specific expression of shHif2a embedded in the miR-E scaffold in the 3’UTR of mCherry. Sequences of shRNAs are provided in Fig. S1 and supplemental data SD1. CC: choriocapillaris, BM: Bruch’s membrane, RPE: retinal pigment epithelium, PS: photoreceptor segments, ONL: outer nuclear layer.

Article Snippet: shRNAmiR DNA fragments containing shHif1a , shHif2a or shCtrl sequences were designed using the miR-E backbone following the recommendation by Fellmann ( ) (Fig. S1A-C) and produced by Bio-Synthesis (Bio-Synthesis Inc. TX, USA).

Techniques: Expressing, Stable Transfection, Comparison, Western Blot, Construct, Control, shRNA, Sequencing, Membrane

Journal: OncoTargets and therapy

Article Title: MicroRNA-145 inhibits the activation of the mTOR signaling pathway to suppress the proliferation and invasion of invasive pituitary adenoma cells by targeting AKT3 in vivo and in vitro

doi: 10.2147/OTT.S118391

Figure Lengend Snippet: Oligodeoxynucleotide sequence of DNA and RNA

Article Snippet: The entire DNA sequence synthesis was performed by Shanghai GenePharma, Co., Ltd (Shanghai, People’s Republic of China; ).

Techniques: Sequencing