A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene
Abstract Background Bioinformatic genome surveys indicate that self-cleaving ribonucleic acids (ribozymes) appear to be widespread among all domains of life, although the functions of only a small number have been validated by biochemical methods. Alternatively, cell-based reporter gene assays can b...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-03-01
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| Series: | BMC Molecular and Cell Biology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12860-021-00357-7 |
| _version_ | 1818741974863183872 |
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| author | Aiyada Aroonsri Jindaporn Kongsee Jeremy David Gunawan Daniel Abidin Aubry Philip James Shaw |
| author_facet | Aiyada Aroonsri Jindaporn Kongsee Jeremy David Gunawan Daniel Abidin Aubry Philip James Shaw |
| author_sort | Aiyada Aroonsri |
| collection | DOAJ |
| description | Abstract Background Bioinformatic genome surveys indicate that self-cleaving ribonucleic acids (ribozymes) appear to be widespread among all domains of life, although the functions of only a small number have been validated by biochemical methods. Alternatively, cell-based reporter gene assays can be used to validate ribozyme function. However, reporter activity can be confounded by phenomena unrelated to ribozyme-mediated cleavage of RNA. Results We established a ribozyme reporter system in Escherichia coli in which a significant reduction of reporter activity is manifest when an active ribozyme sequence is fused to the reporter gene and the expression of a foreign Bacillus subtilis RNaseJ1 5′ exonuclease is induced from a chromosomally-integrated gene in the same cell. Conclusions The reporter system could be useful for validating ribozyme function in candidate sequences identified from bioinformatics. |
| first_indexed | 2024-12-18T02:05:09Z |
| format | Article |
| id | doaj.art-dc0dc7bb38f34a7b835a7fe757b4b658 |
| institution | Directory Open Access Journal |
| issn | 2661-8850 |
| language | English |
| last_indexed | 2024-12-18T02:05:09Z |
| publishDate | 2021-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj.art-dc0dc7bb38f34a7b835a7fe757b4b6582022-12-21T21:24:37ZengBMCBMC Molecular and Cell Biology2661-88502021-03-0122111210.1186/s12860-021-00357-7A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease geneAiyada Aroonsri0Jindaporn Kongsee1Jeremy David Gunawan2Daniel Abidin Aubry3Philip James Shaw4National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)School of Life Science, Indonesia International Institute for Life Sciences (i3L)School of Life Science, Indonesia International Institute for Life Sciences (i3L)National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA)Abstract Background Bioinformatic genome surveys indicate that self-cleaving ribonucleic acids (ribozymes) appear to be widespread among all domains of life, although the functions of only a small number have been validated by biochemical methods. Alternatively, cell-based reporter gene assays can be used to validate ribozyme function. However, reporter activity can be confounded by phenomena unrelated to ribozyme-mediated cleavage of RNA. Results We established a ribozyme reporter system in Escherichia coli in which a significant reduction of reporter activity is manifest when an active ribozyme sequence is fused to the reporter gene and the expression of a foreign Bacillus subtilis RNaseJ1 5′ exonuclease is induced from a chromosomally-integrated gene in the same cell. Conclusions The reporter system could be useful for validating ribozyme function in candidate sequences identified from bioinformatics.https://doi.org/10.1186/s12860-021-00357-7RibozymeE. coli cell-based systemglmS riboswitchHammer-head ribozymeRNaseJ1Reporter system |
| spellingShingle | Aiyada Aroonsri Jindaporn Kongsee Jeremy David Gunawan Daniel Abidin Aubry Philip James Shaw A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene BMC Molecular and Cell Biology Ribozyme E. coli cell-based system glmS riboswitch Hammer-head ribozyme RNaseJ1 Reporter system |
| title | A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene |
| title_full | A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene |
| title_fullStr | A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene |
| title_full_unstemmed | A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene |
| title_short | A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene |
| title_sort | cell based ribozyme reporter system employing a chromosomally integrated 5 exonuclease gene |
| topic | Ribozyme E. coli cell-based system glmS riboswitch Hammer-head ribozyme RNaseJ1 Reporter system |
| url | https://doi.org/10.1186/s12860-021-00357-7 |
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