Small Extracellular Vesicles, or EVs, which measure between 30 and 150 nm in diameter (including exosomes), are naturally released by most living cells. They play a significant role in both physiological and pathological processes, enabling the transfer of information from donor cells to recipient cells. These vesicles carry a variety of cargoes, such as proteins, RNAs, lipids, and DNA, which can be taken up by other cells.
The bioengineering of EVs is facilitated by Ciloa technology, which employs an active sorting mechanism to incorporate any desired protein into the EVs produced by eukaryotic cells, utilizing a proprietary Pilot Peptide (PP) that is patented. This PP interacts with the ESCRT machinery to effectively sort the protein of interest into the EVs. A synthetic DNA sequence, optimized for coding the target protein, is linked to the PP, and the resulting chimeric gene is cloned into Ciloa's proprietary vector. This vector allows for the transcription of mRNA that encodes the chimeric protein in eukaryotic cells. The stable transfection of HEK293T cells results in the expression of the target protein within the EVs.
Fully native proteins conformation
without being hindered by a grafting on an EV protein (e.g. CD63)
Proteins embedded at the EVs surface or in the lumen
Cargo or membrane proteins
Functional hetero-oligomers of membrane protein complexes can be sorted on EVs.
(e.g. hetero-octamer of an ion channel)
Not interfere with regular EVs proteins
(e.g. tetraspanins)
We offer 3 types of Small Extracellular Vesicles
Standard EVs
Standard EVs are native EVs, produced by non-modified cells. These EVs are interesting tools that can be used for data normalisation, EV instrument calibration or as a control, either positive or negative, in EV quality controls, EV characterisation, functional assays, etc.
Fluorescent EVs
Labelled EVs bioengineered with internal cargoes are customised using Ciloa technology which does not overexpress and has no interaction with regular EV surface proteins like tetraspanins. These EVs can be used for data normalisation, purification process internal standard, instrument calibration and cell fluorescence imaging.
- mCerulean EVs
- EGFP EVs
- CyOFP1 EVs
Bioengineered EVs
EVs bioengineered with fully native membrane proteins can harbour any kind of protein like G-protein coupled receptors (GPCR), kinase receptors, ion channels or soluble proteins, fused to a transmembrane domain to anchor them in the EV membrane. They can be use to study molecular interactions with ligands, specific antibodies or small molecules.
- ALK-2 EVs
- ALK-4 EVs
- CCR-10 EVs
- CCR-7 EVs
- MISRII
Specifications
Particule size (nano-flow cytometry) : Median size of 60-70 nm
Source : HEK293T
Buffer : PBS 1X-BSA 10µg/mL
Lifetime: up to 12 months (Standard EVs), up to 6 months (Fluorescent and Bioengineered EVs)
Storage: Store sterile at +4°C
Standard EVs
Standard EVs are native EVs, produced by non-modified cells.
EGFP EVs
EGFP EVs are bioengineered EVs, loaded with EGFP, enhanced green fluorescent protein in their lumen.
mCerulean EVs
mCerulean EVs are bioengineered EVs, loaded with mCerulean cyan fluorescent protein in their lumen.
CyOFP1 EVs
CyOFP1 EVs are bioengineered EVs, loaded with CyOFP1 cyan-excitable orange fluorescent protein in their lumen.
CCR-10 EVs
CCR-10 EVs are bioengineered EVs, harbouring human CCR-10 (human C-C chemokine receptor type 10) GPCR at their surfac
CCR-7 EVs
CCR-7 EVs are bioengineered EVs, harbouring human CCR-7 (human C-C chemokine receptor type ) GPCR at their surface.
ALK-2 EVs
ALK-2 EVs are bioengineered EVs, harbouring human ALK-2 receptor (human Activin receptor-like kinase 2) at their surface.
ALK-4 EVs
ALK-4 EVs are bioengineered EVs, harbouring human ALK-4 receptor (human Activin receptor-like kinase ) at their surface.
MISRII EVs
MISRII EVs are bioengineered EVs, harbouring human MISRII receptor (Muellerian inhibiting substance type II receptor) at their surface
Everspark technology has been intially developed by Karine Monier, Arnaud Favier and Christophe Place and and published in Scientific Reports:
Provost, A., Rousset, C., Bourdon, L. et al. Innovative particle standards and long-lived imaging for 2D and 3D dSTORM. Sci Rep 9, 17967 (2019). https://doi.org/10.1038/s41598-019-53528-0
Publications:
Nanoscale engagement and clusterization of Programmed death ligand 1 (PD-L1) in the membrane lipid rafts of Non-Small Cell Lung Cancer cells
Martina Ruglioni, Simone Civita, Tiziano Salvadori, Sofia Cristiani, Vittoria Carnicelli, Serena Barachini, Iacopo Petrini, Irene Nepita, Marco Castello, Alberto Diaspro, Paolo Bianchini, Barbara Storti, Ranieri Bizzarri, Stefano Fogli and Romano Danesi bioRxiv 2022.08.09.503318; doi: https://doi.org/10.1101/2022.08.09.503318
HIV-1 diverts actin debranching mechanisms for particle assembly and release in CD4 T lymphocytes
Rayane Dibsy, Erwan Bremaud, Johnson Mak, Cyril Favard, Delphine Muriaux
BioRxiv, December 16, 2022. doi. 10.1101/2022.12.15.520580
Fluorescent Polymer-AS1411-Aptamer Probe for dSTORM Super-Resolution Imaging of Endogenous Nucleolin
Fabre L, Rousset C, Monier K, Da Cruz-Boisson F, Bouvet P, Charreyre MT, Delair T, Fleury E, Favier A. Biomacromolecules. 2022 May 12. doi: 10.1021/acs.biomac.1c01706. PMID: 35549176
Comparative analysis of ChAdOx1 nCoV-19 and Ad26.COV2.S SARS-CoV-2 vector vaccines.
Michalik S, Siegerist F, Palankar R, Franzke K, Schindler M, Reder A, Seifert U, Cammann C, Wesche J, Steil L, Hentschker C, Gesell-Salazar M, Reisinger E, Beer M, Endlich N, Greinacher A, Völker U. Haematologica. 2022 Apr 1;107(4):947-957. doi: 10.3324/haematol.2021.280154. PMID: 35045692
Metabolic biorthogonal labeling and dSTORM imaging of peptidoglycan synthesis in Streptococcus pneumoniae
Jennyfer Trouve, Oleksandr Glushonkov and Cecile Morlot
Star Protocols, December 13, 2021. doi: 10.1016/j.xpro.2021.101006
Insights in ChAdOx1 nCoV-19 vaccine-induced immune thrombotic thrombocytopenia.
Greinacher A, Selleng K, Palankar R, Wesche J, Handtke S, Wolff M, Aurich K, Lalk M, Methling K, Völker U, Hentschker C, Michalik S, Steil L, Reder A, Schönborn L, Beer M, Franzke K, Büttner A, Fehse B, Stavrou EX, Rangaswamy C, Mailer RK, Englert H, Frye M, Thiele T, Kochanek S, Krutzke L, Siegerist F, Endlich N, Warkentin TE, Renné T.
Blood. 2021 Dec 2;138(22):2256-2268. doi: 10.1182/blood.2021013231. PMID: 34587242
Superresolution Microscopy of Drosophila Indirect Flight Muscle Sarcomeres.
Szikora S, Novák T, Gajdos T, Erdélyi M, Mihály J. Bio Protoc. 2020 Jun 20;10(12):e3654. doi: 10.21769/BioProtoc.3654. eCollection 2020 Jun 20. PMID: 33659324