Strains

From Kelly Bower, Porter Lab, University of Minnesota, February 2016

Gui L, Song K, Tritschler D, Bower R, Yan S, Dai A, Augspurger K, Sakizadeh J, Grzemska M, Ni T, Porter ME, Nicastro D. Scaffold subunits support associated subunit assembly in the Chlamydomonas ciliary nexin-dynein regulatory complex. Proc Natl Acad Sci U S A. 2019 Nov 12;116(46):23152-23162. doi: 10.1073/pnas.1910960116. Epub 2019 Oct 28. PMID: 31659045; PMCID: PMC6859327.

From Wenqiang Yang, Grossman lab, Carnegie Institution for Science – Department of Plant Biology, February 2016

The fdx5 mutant is disrupted in the second exon by a cassette conferring resistance to paromomycin. It was generated in the CC-4425 (D66+) WT background and was backcrossed 4 times to either CC-124 (mt-) or CC-125 (mt+).

The fdx5 mutant cannot grow in the dark, so it should be maintained on TAP medium in the light. Very low light (> 5 µmol photons m-2 s-1) is sufficient for growth of the fdx5 mutant. It is also resistant to paromomycin.

FDX5 – Cre17.g700950; g16996

Yang W, Wittkopp TM, Li X, Warakanont J, Dubini A, Kim RG, Nowack ECM, Catalanotti C, Aksoy M, Mackinder LCM, Page MD, D’Adamo S, Heinnickel M, Johnson X, Richaud P, Alric J, Boehm M, Jonikas M, Benning C, Merchant SS, Posewitz MC, Grossman AR (2015) Critical role of Chlamydomonas reinhardtii Ferredoxin-5 in maintaining membrane structure and dark metabolism. Proc. Natl. Acad. Sci. USA. 112:14978-14983.

  • Locus:
  • FDX5
  • Chromosome:
  • 17

From Wenqiang Yang, Grossman lab, Carnegie Institution for Science – Department of Plant Biology, February 2016

The fdx5 mutant (CC-5209) is disrupted in the second exon by a cassette conferring resistance to paromomycin. The fdx5-FDX5 complemented strain was generated by transforming FDX5 cDNA (along with the ble resistance gene) into the fdx5 mutant and selecting for growth in the dark on TAP medium supplemented with paromomycin and Zeocin.

The fdx5 mutant cannot grow in the dark, so it should be maintained on TAP medium in the light. The fdx5-FDX5 complemented strain can grow in the light and in the dark, and can be selected for by growth in the presence of paromomycin and Zeocin.

Yang W, Wittkopp TM, Li X, Warakanont J, Dubini A, Kim RG, Nowack ECM, Catalanotti C, Aksoy M, Mackinder LCM, Page MD, D’Adamo S, Heinnickel M, Johnson X, Richaud P, Alric J, Boehm M, Jonikas M, Benning C, Merchant SS, Posewitz MC, Grossman AR (2015) Critical role of Chlamydomonas reinhardtii Ferredoxin-5 in maintaining membrane structure and dark metabolism. Proc. Natl. Acad. Sci. USA. 112:14978-14983.

  • Locus:
  • FDX5
  • Chromosome:
  • 17

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Stable transgenic line derived from CC-1883 in which the plasmid ni-amiRNA-PSY (pAV2) was introduced in order to express an artificial miRNA to silence PSY in an inducible manner. For further details, please see Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Stable transgenic line derived from CC-1883 in which the plasmid ni-amiRNA-PSY (pAV2) was introduced in order to express an artificial miRNA to silence PSY in an inducible manner. For further details, please see Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in the gene encoding AGO3. For further details, please see the description of “GROUP I” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in a non-yet-mapped genomic region. For further details, please see the description of “GROUP III” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • HEN1 (RME3)
  • Chromosome:
  • 3

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in the gene encoding AGO3. For further details, please see the description of “GROUP I” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in the gene encoding DCL3. For further details, please see the description of “GROUP II” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line E9-3, in which an insertional mutagen is inserted in the gene encoding DCL3. For further details, please see the description of “GROUP II” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line E9-3, in which an insertional mutagen is inserted in the gene encoding AGO3. For further details, please see the description of “GROUP II” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in a non-yet-mapped genomic region. For further details, please see the description of “GROUP IV” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line E9-3, in which an insertional mutagen is inserted in the gene encoding DCL3. For further details, please see the description of “GROUP II” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in a non-yet-mapped genomic region. For further details, please see the description of “GROUP IV” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line A4-1, in which an insertional mutagen is inserted in a non-yet-mapped genomic region. For further details, please see the description of “GROUP V” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mutant line that derives from ni-amiRNA-PSY line E9-3, in which an insertional mutagen is inserted in the gene encoding AGO3. For further details, please see the description of “GROUP II” mutants in Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut51 complemented with a BAC (clone A6) that contains the genome sequence encoding the wild type DCL3. For further details, please see Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut51 complemented with a BAC (clone M20) that contains the genome sequence encoding the wild type DCL3. For further details, please see Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut51 complemented with a control plasmid (pALM32) that contains no DCL3 genomic region (negative control of complementation). For further details, please see Valli et al., 2016, Genome Research.

Valli AA, Santos BA, Hnatova S, Bassett AR, Molnar A, Chung BY, Baulcombe DC (2016) Most microRNAs in the single-cell alga Chlamydomonas reinhardtii are produced by Dicer-like 3-mediated cleavage of introns and untranslated regions of coding RNAs. Genome Res. 26:519-29

  • Locus:
  • DCL3
  • Chromosome:
  • 7

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut25 complemented with a plasmid (clone 2) that contains the genome sequence encoding the wild type AGO3. Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut25 complemented with a plasmid (clone 5) that contains the genome sequence encoding the wild type AGO3. Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut33 complemented with a plasmid (clone 2) that contains the genome sequence encoding the wild type AGO3. Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut33 complemented with a plasmid (clone 5) that contains the genome sequence encoding the wild type AGO3. Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut25 complemented with a plasmid (pHyg3) that contains no AGO3 genomic region (negative control of complementation). Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

From Adrian Valli, David Baulcombe lab, University of Cambridge-UK, March 2016

Mut33 complemented with a plasmid (pHyg3) that contains no AGO3 genomic region (negative control of complementation). Valli et al., unpublished results.

Valli et al., unpublished results

  • Locus:
  • AGO3
  • Chromosome:
  • 16

Hung CH, Kanehara K, Nakamura Y. (2016) Isolation and characterization of a mutant defective in triacylglycerol accumulation in nitrogen-starved Chlamydomonas reinhardtii. Biochim Biophys Acta. Apr 7.

  • Locus:
  • BGAL1
  • Chromosome:
  • 2

From Mary Porter, U of MN, April 2016

fla11 mt- (CC-1920) transformed with carboxyl terminally tagged GFP  d1blic by transformation with pSI103  (paromomycin selection)

Unpublished

From Iftach Yacoby, Tel Aviv University, April 2016

Random insertion of the gemonic part of HydEF to rescue the hydEF1 mutant (CC-5128) using the plasmid pSP124s_bleR_HYDEF_KpnI_ins_ (13118bp). Hydrogen production was validated. This was a repeated rescue using the original plasmid from Posewitz et al.

Posewitz MC, King PW, Smolinski SL, Zhang L, Seibert M, Ghirardi ML (2004) Discovery of two novel radical S-adenosylmethionine proteins required for the assembly of an active [Fe] hydrogenase. J Biol Chem. 279:25711-20

  • Locus:
  • HYDEF1
  • Chromosome:
  • 6

From Junya Awata, George Witman lab, University of Massachusetts Medical School, May 2016

The nphp4/drc3 strain, B1179 (nphp4-1, drc3-1, arg7, ARG7, mt+) was first generated by insertional mutagenesis of H11 (arg7, mt+) with the pARG7.8 plasmid and crossed sequentially to the wild-type strains CC-124 and 137c. This drc3 strain was then created by transformation of the crossed B1179 with NPHP4.
Mutant cell lacks first 8 out of 10 exons in DRC3 and has defects in swimming speed and flagellar waveform.

Awata J, Song K, Lin J, King SM, Sanderson MJ, Nicastro D, Witman GB (2015) DRC3 connects the N-DRC to dynein g to regulate flagellar waveform. Mol Biol Cell. 26:2788-2800

  • Locus:
  • DRC3 (FAP134)
  • Chromosome:
  • 12

From Junya Awata, George Witman lab, University of Massachusetts Medical School, May 2016

This double mutant was created by crossing CC-5238 drc3-1 mt+ [D4] to CC-613 pf14 mt-. This strain has the same phenotype as pf14.

Awata J, Song K, Lin J, King SM, Sanderson MJ, Nicastro D, Witman GB (2015) DRC3 connects the N-DRC to dynein g to regulate flagellar waveform. Mol Biol Cell. 26:2788-2800

  • Locus:
  • DRC3 (FAP134), PF14 [RSP3]
  • Chromosome:
  • 12, 6