Descrizione del progetto
Cellule sintetiche mediante RNA Origami
Le cellule sintetiche, note anche come cellule artificiali o protocellule, sono strutture create in laboratorio che imitano alcune proprietà e funzioni delle cellule naturali. Esse hanno suscitato grande interesse per la loro applicazione nella somministrazione di farmaci, nelle biotecnologie e nel rilevamento ambientale, oltre che per il progresso della ricerca biomedica. Finanziato dal Consiglio europeo della ricerca, il progetto ENSYNC intende costruire una cellula sintetica incapsulando biomolecole funzionali all’interno di vescicole lipidiche. L’obiettivo è quello di ottenere una cellula sintetica autoreplicante e in evoluzione attraverso strutture programmabili di origami di RNA, che controllano l’evoluzione e sono funzionalizzate a svolgere diversi compiti. Oltre a comprendere i processi evolutivi, il progetto genererà strumenti basati sugli origami di RNA per varie applicazioni.
Obiettivo
Can we construct a cell from non-living matter? In search for answers, bottom-up synthetic biology has successfully encapsulated functional sets of biomolecules inside lipid vesicles, yet a “living” synthetic cell remains unattained. ENSYNC aims for a prototype of a synthetic cell that encompasses a fundamental characteristic of life, namely evolution. My past work shows that DNA origami can achieve custom-engineered synthetic cellular parts, but the mere encapsulation of preformed parts conflicts with the vision of a self-replicating and evolving synthetic cell. I here propose to produce and to replicate functional RNA origami structures inside of lipid vesicles (GUVs) by co-transcriptional folding from a DNA template. First, I will genetically encode an RNA nanopore and RNA origami structure which induces GUV division. The DNA template (“genotype”) will determine the GUVs’ permeability and their division rate (“phenotype”). This genotype-phenotype mapping is the basis for directed evolution of the rationally engineered RNA origami structures in the second step. In particular, I will aim for efficient GUV division in repeated cycles of genetic diversification and selection. In the third step, I will implement multiple growth and division cycles to enable continuous directed evolution. This will be achieved by system-level integration and laboratory automation of the directed evolution pipeline to iteratively reduce researcher intervention. Depending on externally applied selection pressures, continuous evolution will inevitably lead to the dominance of highly proliferating synthetic cells in mixed populations. ENSYNC provides fundamental insights into evolutionary processes as well as applicable RNA origami-based tools for nanopore sensing and as genetically encoded biophysical probes in cell biology. Overall, ENSYNC pushes the boundaries of bottom-up synthetic biology to the point where synthetic cells can be evolved towards a distinct goal in biotechnology.
Campo scientifico
Parole chiave
Programma(i)
- HORIZON.1.1 - European Research Council (ERC) Main Programme
Argomento(i)
Meccanismo di finanziamento
HORIZON-ERC - HORIZON ERC GrantsIstituzione ospitante
69117 Heidelberg
Germania