AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization.
Immunofluorescence is highly dependent on antibody-antigen interactions for accurate visualization of proteins and other biomolecules within cells. However, obtaining antibodies with high specificity and affinity for their target proteins can be challenging, especially for targets that are complex o...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2024-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0316359 |
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| _version_ | 1841555503617933312 |
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| author | Crystal Chia Yin Ling Melissa Jane Fullwood |
| author_facet | Crystal Chia Yin Ling Melissa Jane Fullwood |
| author_sort | Crystal Chia Yin Ling |
| collection | DOAJ |
| description | Immunofluorescence is highly dependent on antibody-antigen interactions for accurate visualization of proteins and other biomolecules within cells. However, obtaining antibodies with high specificity and affinity for their target proteins can be challenging, especially for targets that are complex or naturally present at low levels. Therefore, we developed AptaFluorescence, a protocol that utilizes fluorescently labeled aptamers for in vitro biomolecule visualization. Aptamers are single-stranded nucleic acid molecules that fold into three-dimensional structures to bind biomolecules with high specificity. AptaFluorescence targeting the c-MYC protein was evaluated in doxycycline-inducible c-MYC U2OS cells. AptaFluorescence signals were more distinct compared to the diffuse immunofluorescence signals. AptaFluorescence also reliably differentiated doxycycline-treated cells from untreated cells. In conclusion, AptaFluorescence is a novel, easy to perform, aptamer-based protocol that will have broad applicability across various biological endeavours for visualizing biomolecules, especially in cases where antibodies with high affinity and specificity for their target proteins are lacking. |
| format | Article |
| id | doaj-art-061f4d186f32446f9bd0c589139f4bb7 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-061f4d186f32446f9bd0c589139f4bb72025-01-08T05:32:21ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-011912e031635910.1371/journal.pone.0316359AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization.Crystal Chia Yin LingMelissa Jane FullwoodImmunofluorescence is highly dependent on antibody-antigen interactions for accurate visualization of proteins and other biomolecules within cells. However, obtaining antibodies with high specificity and affinity for their target proteins can be challenging, especially for targets that are complex or naturally present at low levels. Therefore, we developed AptaFluorescence, a protocol that utilizes fluorescently labeled aptamers for in vitro biomolecule visualization. Aptamers are single-stranded nucleic acid molecules that fold into three-dimensional structures to bind biomolecules with high specificity. AptaFluorescence targeting the c-MYC protein was evaluated in doxycycline-inducible c-MYC U2OS cells. AptaFluorescence signals were more distinct compared to the diffuse immunofluorescence signals. AptaFluorescence also reliably differentiated doxycycline-treated cells from untreated cells. In conclusion, AptaFluorescence is a novel, easy to perform, aptamer-based protocol that will have broad applicability across various biological endeavours for visualizing biomolecules, especially in cases where antibodies with high affinity and specificity for their target proteins are lacking.https://doi.org/10.1371/journal.pone.0316359 |
| spellingShingle | Crystal Chia Yin Ling Melissa Jane Fullwood AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. PLoS ONE |
| title | AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. |
| title_full | AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. |
| title_fullStr | AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. |
| title_full_unstemmed | AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. |
| title_short | AptaFluorescence: An aptamer-based fluorescent imaging protocol for biomolecule visualization. |
| title_sort | aptafluorescence an aptamer based fluorescent imaging protocol for biomolecule visualization |
| url | https://doi.org/10.1371/journal.pone.0316359 |
| work_keys_str_mv | AT crystalchiayinling aptafluorescenceanaptamerbasedfluorescentimagingprotocolforbiomoleculevisualization AT melissajanefullwood aptafluorescenceanaptamerbasedfluorescentimagingprotocolforbiomoleculevisualization |