Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi]
Self-processing hairpin ribozymes have been synthesized from promoterless singlestranded DNA circles (73 nt) within mammalian cells. Following lipid-mediated transient transfection, DNA circles were efficiently internalized by mouse L cells (OST7–1) that stably express T7 RNA polymerase confining...
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| Format: | Article |
| Language: | English |
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De Gruyter
2006-03-01
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| Series: | Türk Biyokimya Dergisi |
| Subjects: | |
| Online Access: | http://www.turkjbiochem.com/2006/002_012.pdf |
| _version_ | 1828032112084647936 |
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| author | Attila A. Seyhan |
| author_facet | Attila A. Seyhan |
| author_sort | Attila A. Seyhan |
| collection | DOAJ |
| description | Self-processing hairpin ribozymes have been synthesized from promoterless singlestranded DNA circles (73 nt) within mammalian cells. Following lipid-mediated transient transfection, DNA circles were efficiently internalized by mouse L cells (OST7–1) that stably express T7 RNA polymerase confining it to the cytoplasm. Cellular uptake of circular DNA templates and intracellular accumulation of ribozyme RNA transcripts from these DNA circles were progressive, both peaking at 24 h after transfection. Intracellular transcription generated RNA concatemers accumulating to a level of ~100 copies per cell. Transcription appears to be independent of specific promoter sequences but depends on T7 RNA polymerase. The data presented here may support the hypothesis that single stranded bubble regions within duplex DNA can serve as de novo initiation sites for RNA transcription not only in vitro but also in the cytoplasm of mammalian cells. These results may provide a model for the rolling circle transcription of small circular nucleic acids in mammalian cells. |
| first_indexed | 2024-04-10T14:59:59Z |
| format | Article |
| id | doaj.art-887677e0d0894a6b980c8dd7497b3fb1 |
| institution | Directory Open Access Journal |
| issn | 0250-4685 1303-829X |
| language | English |
| last_indexed | 2024-04-10T14:59:59Z |
| publishDate | 2006-03-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Türk Biyokimya Dergisi |
| spelling | doaj.art-887677e0d0894a6b980c8dd7497b3fb12023-02-15T16:07:06ZengDe GruyterTürk Biyokimya Dergisi0250-46851303-829X2006-03-01311212Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi]Attila A. SeyhanSelf-processing hairpin ribozymes have been synthesized from promoterless singlestranded DNA circles (73 nt) within mammalian cells. Following lipid-mediated transient transfection, DNA circles were efficiently internalized by mouse L cells (OST7–1) that stably express T7 RNA polymerase confining it to the cytoplasm. Cellular uptake of circular DNA templates and intracellular accumulation of ribozyme RNA transcripts from these DNA circles were progressive, both peaking at 24 h after transfection. Intracellular transcription generated RNA concatemers accumulating to a level of ~100 copies per cell. Transcription appears to be independent of specific promoter sequences but depends on T7 RNA polymerase. The data presented here may support the hypothesis that single stranded bubble regions within duplex DNA can serve as de novo initiation sites for RNA transcription not only in vitro but also in the cytoplasm of mammalian cells. These results may provide a model for the rolling circle transcription of small circular nucleic acids in mammalian cells.http://www.turkjbiochem.com/2006/002_012.pdfT7 RNA polymeraseRNA polymerase IIrTth DNA polymeraserolling circle transcriptionmultimeric RNAsribozyme catalysiscircular DNA |
| spellingShingle | Attila A. Seyhan Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] Türk Biyokimya Dergisi T7 RNA polymerase RNA polymerase II rTth DNA polymerase rolling circle transcription multimeric RNAs ribozyme catalysis circular DNA |
| title | Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] |
| title_full | Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] |
| title_fullStr | Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] |
| title_full_unstemmed | Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] |
| title_short | Rolling circle transcription of ribozymes from promoterless single-stranded DNA nanocircles in mammalian cells [Ribozimlerin, promotorsuz tek-iplikli DNA çemberlerinin rolling circle transkripsiyonu ile memeli hücrelerinde üretimi] |
| title_sort | rolling circle transcription of ribozymes from promoterless single stranded dna nanocircles in mammalian cells ribozimlerin promotorsuz tek iplikli dna cemberlerinin rolling circle transkripsiyonu ile memeli hucrelerinde uretimi |
| topic | T7 RNA polymerase RNA polymerase II rTth DNA polymerase rolling circle transcription multimeric RNAs ribozyme catalysis circular DNA |
| url | http://www.turkjbiochem.com/2006/002_012.pdf |
| work_keys_str_mv | AT attilaaseyhan rollingcircletranscriptionofribozymesfrompromoterlesssinglestrandeddnananocirclesinmammaliancellsribozimlerinpromotorsuztekipliklidnacemberlerininrollingcircletranskripsiyonuilememelihucrelerindeuretimi |