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Organelle trafficking of chimeric ribozymes and genetic manipulation of mitochondria Public Deposited

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  • With the expansion of the RNA world, antisense strategies have become widespread to manipulate nuclear gene expression but organelle genetic systems have remained aside. The present work opens the field to mitochondria. We demonstrate that customized RNAs expressed from a nuclear transgene and driven by a transfer RNA-like (tRNA-like) moiety are taken up by mitochondria in plant cells. The process appears to follow the natural tRNA import specificity, suggesting that translocation indeed occurs through the regular tRNA uptake pathway. Upon validation of the strategy with a reporter sequence, we developed a chimeric catalytic RNA composed of a specially designed trans-cleaving hammerhead ribozyme and a tRNA mimic. Organelle import of the chimeric ribozyme and specific target cleavage within mitochondria were demonstrated in transgenic tobacco cell cultures and Arabidopsis thaliana plants, providing the first directed knockdown of a mitochondrial RNA in a multicellular eukaryote. Further observations point to mitochondrial messenger RNA control mechanisms related to the plant developmental stage and culture conditions. Transformation of mitochondria is only accessible in yeast and in the unicellular alga Chlamydomonas. Based on the widespread tRNA import pathway, our data thus make a breakthrough for direct investigation and manipulation of mitochondrial genetics.
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  • Val, R., Wyszko, E., Valentin, C., Szymanski, M., Cosset, A., Alioua, M., ... Dietrich, A. (2011, November). Organelle trafficking of chimeric ribozymes and genetic manipulation of mitochondria. Nucleic Acids Research, 39(21), 9262-9274. doi:10.1093/nar/gkr580
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  • French Centre National de la Recherche Scientifique (CNRS, UPR2357); Agence Nationale de la Recherche (ANR-09-BLAN-0240-01); Universite´ de Strasbourg (UdS); Polish Academy of Sciences (PAN); exchange programmes (CNRS/PAN cooperation agreement, Polonium, PICS); CNRS/Region Alsace PhD fellowship to R.V. Funding for open access charge: IBMP, CNRS UPR2357.
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  • description.provenance : Submitted by Deanne Bruner (deanne.bruner@oregonstate.edu) on 2012-06-06T23:31:17Z No. of bitstreams: 3 DreherTheoMicrobiologyOrganelleTraffickingChimeric.pdf: 817703 bytes, checksum: 9891576449e2a7285ffe860c149d31ad (MD5) license_rdf: 19965 bytes, checksum: 225316337756db2af069c3edfe03a49f (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
  • description.provenance : Submitted by Deanne Bruner (deanne.bruner@oregonstate.edu) on 2012-06-08T18:30:22Z No. of bitstreams: 3 DreherTheoMicrobiologyOrganelleTraffickingChimeric.pdf: 817703 bytes, checksum: 9891576449e2a7285ffe860c149d31ad (MD5) license_rdf: 19965 bytes, checksum: 225316337756db2af069c3edfe03a49f (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
  • description.provenance : Made available in DSpace on 2012-06-08T18:30:58Z (GMT). No. of bitstreams: 3 DreherTheoMicrobiologyOrganelleTraffickingChimeric.pdf: 817703 bytes, checksum: 9891576449e2a7285ffe860c149d31ad (MD5) license_rdf: 19965 bytes, checksum: 225316337756db2af069c3edfe03a49f (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2011-11
  • description.provenance : Rejected by Sue Kunda(sue.kunda@oregonstate.edu), reason: dfsfsfse on 2012-06-07T17:00:57Z (GMT)
  • description.provenance : Approved for entry into archive by Deanne Bruner(deanne.bruner@oregonstate.edu) on 2012-06-08T18:30:58Z (GMT) No. of bitstreams: 3 DreherTheoMicrobiologyOrganelleTraffickingChimeric.pdf: 817703 bytes, checksum: 9891576449e2a7285ffe860c149d31ad (MD5) license_rdf: 19965 bytes, checksum: 225316337756db2af069c3edfe03a49f (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)

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