Function and anatomy of plant siRNA pools derived from hairpin transgenes
<p>Abstract</p> <p>Background</p> <p>RNA interference results in specific gene silencing by small-interfering RNAs (siRNAs). Synthetic siRNAs provide a powerful tool for manipulating gene expression but high cost suggests that novel siRNA production methods are desirabl...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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BMC
2007-11-01
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| Series: | Plant Methods |
| Online Access: | http://www.plantmethods.com/content/3/1/13 |
| _version_ | 1818964021368324096 |
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| author | Lee Kevin AW Chau Bess L |
| author_facet | Lee Kevin AW Chau Bess L |
| author_sort | Lee Kevin AW |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>RNA interference results in specific gene silencing by small-interfering RNAs (siRNAs). Synthetic siRNAs provide a powerful tool for manipulating gene expression but high cost suggests that novel siRNA production methods are desirable. Strong evolutionary conservation of siRNA structure suggested that siRNAs will retain cross-species function and that transgenic plants expressing heterologous siRNAs might serve as useful siRNA bioreactors. Here we report a detailed evaluation of the above proposition and present evidence regarding structural features of siRNAs extracted from plants.</p> <p>Results</p> <p>Testing the gene silencing capacity of plant-derived siRNAs in mammalian cells proved to be very challenging and required partial siRNA purification and design of a highly sensitive assay. Using the above assay we found that plant-derived siRNAs are ineffective for gene silencing in mammalian cells. Plant-derived siRNAs are almost exclusively double-stranded and most likely comprise a mixture of bona fide siRNAs and aberrant partially complementary duplexes. We also provide indirect evidence that plant-derived siRNAs may contain a hitherto undetected physiological modification, distinct from 3' terminal 2-O-methylation.</p> <p>Conclusion</p> <p>siRNAs produced from plant hairpin transgenes and extracted from plants are ineffective for gene silencing in mammalian cells. Thus our findings establish that a previous claim that transgenic plants offer a cost-effective, scalable and sustainable source of siRNAs is unwarranted. Our results also indicate that the presence of aberrant siRNA duplexes and possibly a plant-specific siRNA modification, compromises the gene silencing capacity of plant-derived siRNAs in mammalian cells.</p> |
| first_indexed | 2024-12-20T12:54:29Z |
| format | Article |
| id | doaj.art-122eef8c66194397875fd3c45a25011d |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-20T12:54:29Z |
| publishDate | 2007-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-122eef8c66194397875fd3c45a25011d2022-12-21T19:40:06ZengBMCPlant Methods1746-48112007-11-01311310.1186/1746-4811-3-13Function and anatomy of plant siRNA pools derived from hairpin transgenesLee Kevin AWChau Bess L<p>Abstract</p> <p>Background</p> <p>RNA interference results in specific gene silencing by small-interfering RNAs (siRNAs). Synthetic siRNAs provide a powerful tool for manipulating gene expression but high cost suggests that novel siRNA production methods are desirable. Strong evolutionary conservation of siRNA structure suggested that siRNAs will retain cross-species function and that transgenic plants expressing heterologous siRNAs might serve as useful siRNA bioreactors. Here we report a detailed evaluation of the above proposition and present evidence regarding structural features of siRNAs extracted from plants.</p> <p>Results</p> <p>Testing the gene silencing capacity of plant-derived siRNAs in mammalian cells proved to be very challenging and required partial siRNA purification and design of a highly sensitive assay. Using the above assay we found that plant-derived siRNAs are ineffective for gene silencing in mammalian cells. Plant-derived siRNAs are almost exclusively double-stranded and most likely comprise a mixture of bona fide siRNAs and aberrant partially complementary duplexes. We also provide indirect evidence that plant-derived siRNAs may contain a hitherto undetected physiological modification, distinct from 3' terminal 2-O-methylation.</p> <p>Conclusion</p> <p>siRNAs produced from plant hairpin transgenes and extracted from plants are ineffective for gene silencing in mammalian cells. Thus our findings establish that a previous claim that transgenic plants offer a cost-effective, scalable and sustainable source of siRNAs is unwarranted. Our results also indicate that the presence of aberrant siRNA duplexes and possibly a plant-specific siRNA modification, compromises the gene silencing capacity of plant-derived siRNAs in mammalian cells.</p>http://www.plantmethods.com/content/3/1/13 |
| spellingShingle | Lee Kevin AW Chau Bess L Function and anatomy of plant siRNA pools derived from hairpin transgenes Plant Methods |
| title | Function and anatomy of plant siRNA pools derived from hairpin transgenes |
| title_full | Function and anatomy of plant siRNA pools derived from hairpin transgenes |
| title_fullStr | Function and anatomy of plant siRNA pools derived from hairpin transgenes |
| title_full_unstemmed | Function and anatomy of plant siRNA pools derived from hairpin transgenes |
| title_short | Function and anatomy of plant siRNA pools derived from hairpin transgenes |
| title_sort | function and anatomy of plant sirna pools derived from hairpin transgenes |
| url | http://www.plantmethods.com/content/3/1/13 |
| work_keys_str_mv | AT leekevinaw functionandanatomyofplantsirnapoolsderivedfromhairpintransgenes AT chaubessl functionandanatomyofplantsirnapoolsderivedfromhairpintransgenes |