Quinine copolymer reporters promote efficient intracellular DNA delivery and illuminate a protein-induced unpackaging mechanism.

Polymeric vehicles that efficiently package and controllably release nucleic acids enable the development of safer and more efficacious strategies in genetic and polynucleotide therapies. Developing delivery platforms that endogenously monitor the molecular interactions, which facilitate binding and release of nucleic acids in cells, would aid in the rational design of more effective vectors for clinical applications. Here, we report the facile synthesis of a copolymer containing quinine and 2-hydroxyethyl acrylate that effectively compacts plasmid DNA (pDNA) through electrostatic binding and intercalation. This polymer system poly(quinine-co-HEA) packages pDNA and shows exceptional cellular internalization, transgene expression, and low cytotoxicity compared to commercial controls for several human cell lines, including HeLa, HEK 293T, K562, and keratinocytes (N/TERTs). Using quinine as an endogenous reporter for pDNA intercalation, Raman imaging revealed that proteins inside cells facilitate the unpackaging of polymer-DNA complexes (polyplexes) and the release of their cargo. Our work showcases the ability of this quinine copolymer reporter to not only facilitate effective gene delivery but also enable diagnostic monitoring of polymer-pDNA binding interactions on the molecular scale via Raman imaging. The use of Raman chemical imaging in the field of gene delivery yields unprecedented insight into the unpackaging behavior of polyplexes in cells and provides a methodology to assess and design more efficient delivery vehicles for gene-based therapies.

Making Olympic lizards: the effects of specialised exercise training on performance.

Exercise training is well known to affect a suite of physiological and performance traits in mammals, but effects of training in other vertebrate tetrapod groups have been inconsistent. We examined performance and physiological differences among green anole lizards (Anolis carolinensis) that were trained for sprinting or endurance, using an increasingly rigorous training regimen over 8 weeks. Lizards trained for endurance had significantly higher post-training endurance capacity compared with the other treatment groups, but groups did not show post-training differences in sprint speed. Although acclimation to the laboratory environment and training explain some of our results, mechanistic explanations for these results correspond with the observed performance differences. After training, endurance-trained lizards had higher haematocrit and larger fast glycolytic muscle fibres. Despite no detectable change in maximal performance of sprint-trained lizards, we detected that they had significantly larger slow oxidative muscle fibre areas compared with the other treatments. Treatment groups did not differ in the proportion of number of fibre types, nor in the mass of most limb muscles or the heart. Our results offer some caveats for investigators conducting training research on non-model organisms and they reveal that muscle plasticity in response to training may be widespread phylogenetically.

Leading edge: emerging drug, cell, and gene therapies for junctional epidermolysis bullosa.

INTRODUCTION: Junctional epidermolysis bullosa (JEB) is a rare inherited genetic disorder with limited treatments beyond palliative care. A major hallmark of JEB is skin blistering caused by functional loss or complete absence of major structural proteins of the skin. Impaired wound healing in patients with JEB gives rise to chronic cutaneous ulcers that require daily care. Wound care and infection control are the current standard of care for this patient population. AREAS COVERED: This review covers research and clinical implementation of emerging drug, cell, and gene therapies for JEB. Current clinical trials use topical drug delivery to manipulate the inflammation and re-epithelialization phases of wound healing or promote premature stop codon readthrough to accelerate chronic wound closure. Allogeneic cell therapies for JEB have been largely unsuccessful, with autologous skin grafting emerging as a reliable method of resolving the cutaneous manifestations of JEB. Genetic correction and transplant of autologous keratinocytes have demonstrated persistent amelioration of chronic wounds in a subset of patients. EXPERT OPINION: Emerging therapies address the cutaneous symptoms of JEB but are unable to attend to systemic manifestations of the disease. Investigations into the molecular mechanism(s) underpinning the failure of systemic allogeneic cell therapies are necessary to expand the range of effective JEB therapies.