An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity
Detection of Ribonucleic acids (RNA) is a critical step in the identification of viral or bacterial infections in humans and animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) remains the gold standard, but clustered regularly interspaced short palindromic repeats linked to a Cas endo...
Saved in:
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-05-01
|
| Series: | Biosensors and Bioelectronics: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590137025000172 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1823856773294981120 |
|---|---|
| author | Pierre Guermonprez Le Tu Anh Louis Renaud Pierre Nioche Eric Krejci Aurélie Alleaume-Butaux Nicolas Battaglini Vu Thi Thu Sébastien Sanaur Benoît Piro |
| author_facet | Pierre Guermonprez Le Tu Anh Louis Renaud Pierre Nioche Eric Krejci Aurélie Alleaume-Butaux Nicolas Battaglini Vu Thi Thu Sébastien Sanaur Benoît Piro |
| author_sort | Pierre Guermonprez |
| collection | DOAJ |
| description | Detection of Ribonucleic acids (RNA) is a critical step in the identification of viral or bacterial infections in humans and animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) remains the gold standard, but clustered regularly interspaced short palindromic repeats linked to a Cas endoribonuclease particle (CRISPR/Cas) have recently revolutionized the recognition step of two RNAs, i.e. the CRISPR-RNA (crRNA) and the target, providing a much better selectivity compared to the naked hybridization on which RT-PCR is based. Here, we combine the high efficiency of the CRISPR/Cas13a system with the transduction and amplification capabilities of an electrolyte-gated graphene field-effect transistor (EGGFET) for the detection of specific RNA sequences. In these devices, fabricated on flexible plastic substrates, the active material (reduced graphene oxide, rGO) is deposited by printing and then functionalized with Au nanoparticles decorated with polyU RNA reporter strands. In this system, the CRISPR/Cas13a complex acts as a catalyst: in the presence of a specific RNA target sequence, the enzymatic function is activated and the polyU RNA reporter strands are cleaved from the Au nanoparticles, inducing a loss of negative charges on the rGO layer. This phenomenon leads to measurable changes in the transfer curve of the transistors. These sensors were tested for the detection of a SARS-CoV-2 RNA sequence and showed a linear response in the range of 10−7 - 102 ng μL−1 with an estimated limit of detection of 10 fM. This work is an important milestone in the development of the next generation of point-of-care RNA sensors. |
| format | Article |
| id | doaj-art-c8b066b823b24e4f9c15aba8efda5d7e |
| institution | Kabale University |
| issn | 2590-1370 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosensors and Bioelectronics: X |
| spelling | doaj-art-c8b066b823b24e4f9c15aba8efda5d7e2025-02-12T05:31:45ZengElsevierBiosensors and Bioelectronics: X2590-13702025-05-0123100590An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activityPierre Guermonprez0Le Tu Anh1Louis Renaud2Pierre Nioche3Eric Krejci4Aurélie Alleaume-Butaux5Nicolas Battaglini6Vu Thi Thu7Sébastien Sanaur8Benoît Piro9Université Paris Cité, ITODYS, CNRS, F-75006, Paris, FranceUniversité Paris Cité, ITODYS, CNRS, F-75006, Paris, France; University of Science and Technology of Hanoi (USTH) and Academy of Science and Technology (VAST), 18 Hoang Quoc Viet, Hanoi, Viet NamUniversité de Lyon, Institut des Nanotechnologies de Lyon INL-UMR5270, Université Lyon 1, F-69622, Villeurbanne, FranceUniversité Paris Cité, Paris, Structural and Molecular Analysis Platform, INSERM US 36 | CNRS UAR 2009, Paris, France; Université Paris Cité, INSERM U1124, Paris, FranceCNRS, Université Paris Cité, ENS Paris Saclay, Centre Borelli UMR 9010, Paris, FranceUniversité Paris Cité, Paris, Structural and Molecular Analysis Platform, INSERM US 36 | CNRS UAR 2009, Paris, France; Université Paris Cité, INSERM U1124, Paris, FranceUniversité Paris Cité, ITODYS, CNRS, F-75006, Paris, FranceUniversity of Science and Technology of Hanoi (USTH) and Academy of Science and Technology (VAST), 18 Hoang Quoc Viet, Hanoi, Viet NamInstitut Mines-Telecom, Mines Saint-Étienne, Department of Flexible Electronics, Gardanne, F-13541, FranceUniversité Paris Cité, ITODYS, CNRS, F-75006, Paris, France; Corresponding author.Detection of Ribonucleic acids (RNA) is a critical step in the identification of viral or bacterial infections in humans and animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) remains the gold standard, but clustered regularly interspaced short palindromic repeats linked to a Cas endoribonuclease particle (CRISPR/Cas) have recently revolutionized the recognition step of two RNAs, i.e. the CRISPR-RNA (crRNA) and the target, providing a much better selectivity compared to the naked hybridization on which RT-PCR is based. Here, we combine the high efficiency of the CRISPR/Cas13a system with the transduction and amplification capabilities of an electrolyte-gated graphene field-effect transistor (EGGFET) for the detection of specific RNA sequences. In these devices, fabricated on flexible plastic substrates, the active material (reduced graphene oxide, rGO) is deposited by printing and then functionalized with Au nanoparticles decorated with polyU RNA reporter strands. In this system, the CRISPR/Cas13a complex acts as a catalyst: in the presence of a specific RNA target sequence, the enzymatic function is activated and the polyU RNA reporter strands are cleaved from the Au nanoparticles, inducing a loss of negative charges on the rGO layer. This phenomenon leads to measurable changes in the transfer curve of the transistors. These sensors were tested for the detection of a SARS-CoV-2 RNA sequence and showed a linear response in the range of 10−7 - 102 ng μL−1 with an estimated limit of detection of 10 fM. This work is an important milestone in the development of the next generation of point-of-care RNA sensors.http://www.sciencedirect.com/science/article/pii/S2590137025000172CRISPR/CasElectrolyte-gated transistorReduced graphene oxideRNA |
| spellingShingle | Pierre Guermonprez Le Tu Anh Louis Renaud Pierre Nioche Eric Krejci Aurélie Alleaume-Butaux Nicolas Battaglini Vu Thi Thu Sébastien Sanaur Benoît Piro An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity Biosensors and Bioelectronics: X CRISPR/Cas Electrolyte-gated transistor Reduced graphene oxide RNA |
| title | An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity |
| title_full | An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity |
| title_fullStr | An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity |
| title_full_unstemmed | An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity |
| title_short | An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity |
| title_sort | electrolyte gated transistor for the monitoring of a crispr cas activity |
| topic | CRISPR/Cas Electrolyte-gated transistor Reduced graphene oxide RNA |
| url | http://www.sciencedirect.com/science/article/pii/S2590137025000172 |
| work_keys_str_mv | AT pierreguermonprez anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT letuanh anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT louisrenaud anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT pierrenioche anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT erickrejci anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT aureliealleaumebutaux anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT nicolasbattaglini anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT vuthithu anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT sebastiensanaur anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT benoitpiro anelectrolytegatedtransistorforthemonitoringofacrisprcasactivity AT pierreguermonprez electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT letuanh electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT louisrenaud electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT pierrenioche electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT erickrejci electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT aureliealleaumebutaux electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT nicolasbattaglini electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT vuthithu electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT sebastiensanaur electrolytegatedtransistorforthemonitoringofacrisprcasactivity AT benoitpiro electrolytegatedtransistorforthemonitoringofacrisprcasactivity |