Oligonucleotides have shifted drug discovery into a new paradigm due to their ability to silence the genes and inhibit protein translation. Importantly, they can drug the un-druggable targets from the conventional small-molecule perspective. Unfortunately, poor cellular permeability and susceptibility to nuclease degradation remain as major hurdles for the development of oligonucleotide therapeutic agents. Studies of safe and effective delivery technique with lipid bioconjugates gains attention to resolve these issues. Our review article summarizes the physicochemical effect of well-studied hydrophobic moieties to enhance the cellular entry of oligonucleotides. The structural impacts of fatty acids, cholesterol, tocopherol, and squalene on cellular internalization and membrane penetration in vitro and in vivo were discussed first. The crucial assays for delivery evaluation within this section were analyzed sequentially. Next, we provided a few successful examples of lipid-conjugated oligonucleotides advanced into clinical studies for treating patients with different medical backgrounds. Finally, we pinpointed current limitations and outlooks in this research field along with opportunities to explore new modifications and efficacy studies.

Feedforward loops (FFLs) are relatively simple network motifs, made of three interacting genes, that have been found in a large number in E. coli and S. cerevisiae. More recently, they have also been discovered in multicellular eukaryotes. FFLs are evolutionary favored motifs because they enable cells to survive critical environmental conditions. Among the eight types of possible FFLs, the so-called coherent 1 and incoherent 1 FFL are the most abundant. The former carries out a sign-sensitive delay in gene expression; the latter is a pulse generator and a response time accelerator. So far, only few synthetic FFLs have been engineered, either in cell-free systems or in vivo. In this work, we review the main experimental works published on FFLs, with particular focus on novel designs for synthetic FFLs. They are, indeed, quite different from the natural ones that arose during the course of evolution.