Author(s):
Rebelo, Catarina ; Reis, Tiago ; Guedes, Joana ; Saraiva, Cláudia ; Rodrigues, Artur Filipe ; Simões, Susana ; Bernardino, Liliana ; Peça, João ; Pinho, Sónia L. C. ; Ferreira, Lino
Date: 2022
Persistent ID: https://hdl.handle.net/10316/101264
Origin: Estudo Geral - Universidade de Coimbra
Project/scholarship:
info:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC/PT;
info:eu-repo/grantAgreement/other///other;
info:eu-repo/grantAgreement/EC/H2020/669088/EU;
info:eu-repo/grantAgreement/EC/H2020/952266/EU;
info:eu-repo/grantAgreement/FCT/OE/PD/PT;
info:eu-repo/grantAgreement/FCT/FARH/SFRH/PT;
info:eu-repo/grantAgreement/EC/H2020/101003413/EU;
Subject(s): Brain; Infrared Rays; Optogenetics; Proteins; Gene Editing; Nanoparticles
Description
Spatial control of gene expression is critical to modulate cellular functions and deconstruct the function of individual genes in biological processes. Light-responsive gene-editing formulations have been recently developed; however, they have shown limited applicability in vivo due to poor tissue penetration, limited cellular transfection and the difficulty in evaluating the activity of the edited cells. Here, we report a formulation composed of upconversion nanoparticles conjugated with Cre recombinase enzyme through a photocleavable linker, and a lysosomotropic agent that facilitates endolysosomal escape. This formulation allows in vitro spatial control in gene editing after activation with near-infrared light. We further demonstrate the potential of this formulation in vivo through three different paradigms: (i) gene editing in neurogenic niches, (ii) gene editing in the ventral tegmental area to facilitate monitoring of edited cells by precise optogenetic control of reward and reinforcement, and (iii) gene editing in a localized brain region via a noninvasive administration route (i.e., intranasal).
L.F. acknowledges the funding by COMPETE2020 and FCT programs [POCI-01-0145- FEDER-016390 (acronym: CANCEL STEM) and PTDC/NAN-MAT/28060/2017 (acronym: BrainEdition), POCI-01-0145-FEDER-016682], Portugal 2020-COMPETE funding through “Programa Operacional Regional do Centro” CENTRO2020 (CENTRO-01-0145-FEDER-000014) and Projects Interreg 2IQBioNeuro (“Impulso de una red de I + ien química biológica para diagnóstico y tratamiento de enfermedades neurológicas”) and NeuroAtlantic (“An Atlantic innovation platform on diagnosis and treatment of neurological diseases and aging”, ref: EAPA_791/2018), and EC projects ERAatUC (Ref:669088) and RESETageing (Ref:952266). C.R. and S.P. acknowledge the FCT fellowships (SFRH/BD/52337/2013; SFRH/BPD/96048/2013). J.P. acknowledges funding from BIAL Foundation (BIAL 266/2016). A.F.R. is grateful to the EU Horizon 2020 programme for funding under grant agreement no. 101003413. The authors would like to thank Claudio Franco and Catarina Fonseca from IMM for the multiphoton microscopy studies.
