Imperial College London

Liliana Brito

Faculty of MedicineNational Heart & Lung Institute

Casual - Student demonstrator - lower rate
 
 
 
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Contact

 

l.brito16

 
 
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Location

 

Royal School of MinesSouth Kensington Campus

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Summary

 

Publications

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2 results found

Hachim D, Zhao J, Bhankharia J, Nuñez-Toldra R, Brito L, Seong H, Becce M, Ouyang L, Grigsby C, Higgins S, Terracciano C, Stevens Met al., 2022, Polysaccharide-polyplex nanofilm coatings enhance nanoneedle-based gene delivery and transfection efficiency, Small, Vol: 18, ISSN: 1613-6810

Non-viral vectors represent versatile and immunologically safer alternatives for nucleic acid delivery. Nanoneedles and high-aspect ratio nanostructures are unconventional but interesting delivery systems, in which delivery is mediated by surface interactions. Herein, nanoneedles are synergistically combined with polysaccharide-polyplex nanofilms and enhanced transfection efficiency is observed, compared to polyplexes in suspension. Different polyplex-polyelectrolyte nanofilm combinations are assessed and it is found that transfection efficiency is enhanced when using polysaccharide-based polyanions, rather than being only specific for hyaluronic acid, as suggested in earlier studies. Moreover, results show that enhanced transfection is not mediated by interactions with the CD44 receptor, previously hypothesized as a major mechanism mediating enhancement via hyaluronate. In cardiac tissue, nanoneedles are shown to increase the transfection efficiency of nanofilms compared to flat substrates; while in vitro, high transfection efficiencies are observed in nanostructures where cells present large interfacing areas with the substrate. The results of this study demonstrate that surface-mediated transfection using this system is efficient and safe, requiring amounts of nucleic acid with an order of magnitude lower than standard culture transfection. These findings expand the spectrum of possible polyelectrolyte combinations that can be used for the development of suitable non-viral vectors for exploration in further clinical trials.

Journal article

Moroz-Omori EV, Satyapertiwi D, Ramel MC, Hogset H, Sunyovszki I, Liu Z, Wojciechowski J, Zhang Y, Grigsby CL, Casimiro Brito L, Bugeon L, Dallman M, Stevens Met al., 2020, Photoswitchable gRNAs for spatiotemporally controlled CRISPR-Cas-based genomic regulation, ACS Central Science, Vol: 6, Pages: 695-703, ISSN: 2374-7943

The recently discovered CRISPR-Cas gene editing system and its derivatives have found numerous applications in fundamental biology research and pharmaceutical sciences. The need for precise external control over the gene editing and regulatory events has driven the development of inducible CRISPR-Cas systems. While most of the light-controllable CRISPR-Cas systems are based on protein engineering, we developed an alternative synthetic approach based on modification of crRNA/tracrRNA duplex (guide RNA or gRNA) with photocaging groups, preventing the gRNA from recognizing its genome target sequence until its deprotection is induced within seconds of illumination. This approach relies on a straightforward solid-phase synthesis of the photocaged gRNAs, with simpler purification and characterization processes in comparison to engineering a light-responsive protein. We have demonstrated the feasibility of photocaging of gRNAs and light-mediated DNA cleavage upon brief exposure to light in vitro. We have achieved light-mediated spatiotemporally resolved gene editing as well as gene activation in cells, whereas photocaged gRNAs showed virtually no detectable gene editing or activation in the absence of light irradiation. Finally, we have applied this system to spatiotemporally control gene editing in zebrafish embryos in vivo, enabling the use of this strategy for developmental biology and tissue engineering applications.

Journal article

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