Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas.

Cerutti, Heriberto; Johnson, Anita; Gillham, Nicholas W.; Boynton, John E.

doi:10.1105/tpc.9.6.925

Cited by 189 publications

(160 citation statements)

References 82 publications

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“…Yet this alga also relies on a DCL1-independent, transcriptional silencing mechanism(s) to control mobile genetic elements. Interestingly, chromatin-mediated repression involving the MUT11 machinery is sensitive to temperature, being much more effective at 17°than at 25° (Cerutti et al 1997b). Conversely, post-transcriptional gene silencing by RNAi, in both invertebrates and higher plants, appears to be more efficient at 25°-29°than at lower temperatures (Fortier and Belote 2000;Szittya et al 2003).…”

Section: Discussionmentioning

confidence: 99%

Diversification of the Core RNA Interference Machinery in Chlamydomonas reinhardtii and the Role of DCL1 in Transposon Silencing

Casas-Mollano¹,

Rohr²,

Kim

et al. 2008

Genetics

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Small RNA-guided gene silencing is an evolutionarily conserved process that operates by a variety of molecular mechanisms. In multicellular eukaryotes, the core components of RNA-mediated silencing have significantly expanded and diversified, resulting in partly distinct pathways for the epigenetic control of gene expression and genomic parasites. In contrast, many unicellular organisms with small nuclear genomes seem to have lost entirely the RNA-silencing machinery or have retained only a basic set of components. We report here that Chlamydomonas reinhardtii, a unicellular eukaryote with a relatively large nuclear genome, has undergone extensive duplication of Dicer and Argonaute polypeptides after the divergence of the green algae and land plant lineages. Chlamydomonas encodes three Dicers and three Argonautes with DICER-LIKE1 (DCL1) and ARGONAUTE1 being more divergent than the other paralogs. Interestingly, DCL1 is uniquely involved in the post-transcriptional silencing of retrotransposons such as TOC1. Moreover, on the basis of the subcellular distribution of TOC1 small RNAs and target transcripts, this pathway most likely operates in the nucleus. However, Chlamydomonas also relies on a DCL1-independent, transcriptional silencing mechanism(s) for the maintenance of transposon repression. Our results suggest that multiple, partly redundant epigenetic processes are involved in preventing transposon mobilization in this green alga.

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Section: Discussionmentioning

confidence: 99%

Diversification of the Core RNA Interference Machinery in Chlamydomonas reinhardtii and the Role of DCL1 in Transposon Silencing

Casas-Mollano¹,

Rohr²,

Kim

et al. 2008

Genetics

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“…Participation of each mechanism is very often dependent on the production of free ends generated by DSBs and the latter is dependent on the availability of processing enzymes typical for a particular mechanism (Sonoda et al 2006). Additionally, when we consider well-known difficulties in heterologous gene expression in Chlamydomonas cells (Cerutti et al 1997), this approach for improving HR in Chlamydomonas is not very promising.…”

Section: Expression Of the Genes Responsible For Hrmentioning

confidence: 99%

Is it possible to improve homologous recombination in Chlamydomonas reinhardtii?

et al. 2008

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Targeted modification of the genome has long been an aim of many geneticists and biotechnologists. Gene targeting is a main molecular tool to examine biological effects of genes in a controlled environment. Effective gene targeting depends on the frequency of homologous recombination that is indispensable for the insertion of foreign DNA into a specific sequence of the genome. The main problem associated with the development of an optimal procedure for gene targeting in a particular organism is the variability of homologous recombination (HR) in different species. Chlamydomonas reinhardtii is an attractive model system for the study of many cellular processes and is also an interesting object for the biotechnology industry. In spite of many advantages of this model system, C. reinhardtii does not readily express heterologous genes and does not allow targeted integration of foreign DNA into its genome easily. This paper compares data obtained from several different experiments designed for improving gene targeting in different organisms and reviews the suitability of particular techniques in C. reinhardtii cells.

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“…Chlamydomonas cells were routinely grown in TAP medium under moderate light conditions (20,26,40). Reactivation of expression of the RbcS2:aadA:RbcS2 transgene was examined by spotting dilutions of cells on TAP plates containing spectinomycin (39). To test whether the insertional mutagen cosegregated with reactivation of transgenic expression, we crossed Mut-9 and Mut-11 to the wild-type strain of opposite mating type, CC-125, and dissected tetrads as previously described (40).…”

Section: Methodsmentioning

confidence: 99%

“…The 11-P[300], 1-P[300], and Mut-11 strains have been previously described (20,26,39). Chlamydomonas cells were routinely grown in TAP medium under moderate light conditions (20,26,40).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

The MUT9p kinase phosphorylates histone H3 threonine 3 and is necessary for heritable epigenetic silencing in Chlamydomonas

Casas-Mollano¹,

Jeong²,

Xu³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

101

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Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas.

Cited by 189 publications

References 82 publications

Diversification of the Core RNA Interference Machinery in Chlamydomonas reinhardtii and the Role of DCL1 in Transposon Silencing

Diversification of the Core RNA Interference Machinery in Chlamydomonas reinhardtii and the Role of DCL1 in Transposon Silencing

Is it possible to improve homologous recombination in Chlamydomonas reinhardtii?

The MUT9p kinase phosphorylates histone H3 threonine 3 and is necessary for heritable epigenetic silencing in Chlamydomonas

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