BackgroundProtein-based Cas9 in vivo gene editing therapeutics have practical limitations owing to their instability and low efficacy. To overcome these obstacles and improve stability, we designed a nanocarrier primarily consisting of lecithin that can efficiently target liver disease and encapsulate complexes of Cas9 with a single-stranded guide RNA (sgRNA) ribonucleoprotein (Cas9-RNP) through polymer fusion self-assembly.ResultsIn this study, we optimized an sgRNA sequence specifically for dipeptidyl peptidase-4 gene (DPP-4) to modulate the function of glucagon-like peptide 1. We then injected our nanocarrier Cas9-RNP complexes directly into type 2 diabetes mellitus (T2DM) db/db mice, which disrupted the expression of DPP-4 gene in T2DM mice with remarkable efficacy. The decline in DPP-4 enzyme activity was also accompanied by normalized blood glucose levels, insulin response, and reduced liver and kidney damage. These outcomes were found to be similar to those of sitagliptin, the current chemical DPP-4 inhibition therapy drug which requires recurrent doses.ConclusionsOur results demonstrate that a nano-liposomal carrier system with therapeutic Cas9-RNP has great potential as a platform to improve genomic editing therapies for human liver diseases.Electronic supplementary materialThe online version of this article (10.1186/s12951-019-0452-8) contains supplementary material, which is available to authorized users.
Real‐time in vivo detection of cancer via attenuation‐based plain X‐ray imaging is proposed to fundamentally overcome the penetration depth limits of current fluorescence‐based imaging techniques. Using cesium lead bromide (CsPbBr3, CPB) quantum dot (QD) scintillators, real‐time X‐ray detection of 5 mm‐sized Panc‐1 cell tumors grown in a mouse is successfully performed. The QDs are rapidly co‐synthesized and double‐encapsulated with silicon dioxide (SiO2) to completely prevent them from being aggregated, decomposed, or released; they are then conjugated with antibodies to target pancreatic cancer. Due to the dramatic X‐ray attenuation, the X‐ray signal from the CPB QDs placed under the 2 cm‐thick tissue is clearly observed, while their fluorescence signal is not detected at all. In in vivo mouse experiments, the injection of a tiny amount (2.8 μg on a QD basis) of the CPB–SiO2@SiO2–Ab nanoparticles gives rise to a bright spot at the location of the tumor. Cell viability assay and histological analysis confirm the biocompatibility and nontoxicity of the nanoparticles.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.