RESUMO
Interest in algae has significantly accelerated with the increasing recognition of their potentially unique role in medical, materials, energy, bioremediation, and synthetic biological research. However, the introduction of tools to study, control, or expand the inner-workings of algae has lagged behind. Here we describe a general molecular method based on guanidinium-rich molecular transporters (GR-MoTrs) for bringing small and large cargos into algal cells. Significantly, this method is shown to work in wild-type algae that have an intact cell wall. Developed using Chlamydomonas reinhardtii, this method is also successful with less studied algae including Neochloris oleoabundans and Scenedesmus dimorphus thus providing a new and versatile tool for algal research.
Assuntos
Bioquímica/métodos , Parede Celular/metabolismo , Chlamydomonas reinhardtii/citologia , Chlamydomonas reinhardtii/metabolismo , Sondas Moleculares/metabolismo , Proteínas/metabolismo , Transporte Biológico , Escuridão , Flagelos/metabolismo , Citometria de Fluxo , Fluoresceína/metabolismo , Microscopia de Fluorescência , TemperaturaRESUMO
The polyanionic nature of oligonucleotides and their enzymatic degradation present challenges for the use of siRNA in research and therapy; among the most notable of these is clinically relevant delivery into cells. To address this problem, we designed and synthesized the first members of a new class of guanidinium-rich amphipathic oligocarbonates that noncovalently complex, deliver, and release siRNA in cells, resulting in robust knockdown of target protein synthesis in vitro as determined using a dual-reporter system. The organocatalytic oligomerization used to synthesize these co-oligomers is step-economical and broadly tunable, affording an exceptionally quick strategy to explore chemical space for optimal siRNA delivery in varied applications. The speed and versatility of this approach and the biodegradability of the designed agents make this an attractive strategy for biological tool development, imaging, diagnostics, and therapeutic applications.
Assuntos
Carbonatos/química , Guanidina/química , Queratinócitos/metabolismo , RNA Interferente Pequeno/metabolismo , Transporte Biológico/efeitos dos fármacos , Carbonatos/síntese química , Carbonatos/toxicidade , Morte Celular/efeitos dos fármacos , Células Cultivadas , Citometria de Fluxo , Técnicas de Transferência de Genes , Genes Reporter/genética , Proteínas de Fluorescência Verde/metabolismo , Guanidina/síntese química , Guanidina/toxicidade , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Luz , Solanum lycopersicum/metabolismo , Microscopia de Fluorescência , RNA Interferente Pequeno/toxicidade , Espalhamento de RadiaçãoRESUMO
Inspired originally by peptides that traverse biological barriers, research on molecular transporters has since identified the key structural requirements that govern cellular entry, leading to new, significantly more effective and more readily available agents. These new drug delivery systems enable or enhance cellular and tissue uptake, can be targeted, and provide numerous additional advantages of significance in imaging, diagnostics and therapy.