Strains

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

Dymek EE, Lefebvre PA, Smith EF (2004) PF15p is the chlamydomonas homologue of the Katanin p80 subunit and is required for assembly of flagellar central microtubules. Eukaryot Cell. 3:870-9

  • Locus:
  • PF15 [KAT2]
  • Chromosome:
  • 3

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

Dymek EE, Lefebvre PA, Smith EF (2004) PF15p is the chlamydomonas homologue of the Katanin p80 subunit and is required for assembly of flagellar central microtubules. Eukaryot Cell. 3:870-9

  • Locus:
  • PF15 [KAT2]
  • Chromosome:
  • 3

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of FAP74

DiPetrillo CG & Smith EF (2010) Pcdp1 is a central apparatus protein that binds Ca(2+)-calmodulin and regulates ciliary motility. J Cell Biol. 189:601-12

  • Locus:
  • FAP74
  • Chromosome:
  • 6

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of FAP74

DiPetrillo CG & Smith EF (2010) Pcdp1 is a central apparatus protein that binds Ca(2+)-calmodulin and regulates ciliary motility. J Cell Biol. 189:601-12

  • Locus:
  • FAP74
  • Chromosome:
  • 6

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of CaM-IP2 (FAP91)

Dymek EE, Heuser T, Nicastro D, Smith EF (2011) The CSC is required for complete radial spoke assembly and wild-type ciliary motility. Mol Biol Cell. 22:2520-31

  • Locus:
  • FAP91
  • Chromosome:
  • 7

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of CaM-IP2 (FAP91)

Dymek EE, Heuser T, Nicastro D, Smith EF (2011) The CSC is required for complete radial spoke assembly and wild-type ciliary motility. Mol Biol Cell. 22:2520-31

  • Locus:
  • FAP91
  • Chromosome:
  • 7

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of CaM-IP3 (FAP61)

Dymek EE, Heuser T, Nicastro D, Smith EF (2011) The CSC is required for complete radial spoke assembly and wild-type ciliary motility. Mol Biol Cell. 22:2520-31

  • Locus:
  • FAP61
  • Chromosome:
  • 3

If you use this mutant for your work, please cite: Li et al. 2019 Nature Genetics.

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

Confirmed no FAP91 by western blot

  • Locus:
  • FAP91
  • Chromosome:
  • 7

If you use this mutant for your work, please cite: Li et al. 2019 Nature Genetics.

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

Confirmed no FAP61 by western blot

  • Locus:
  • FAP61
  • Chromosome:
  • 3

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

pf6-2 (CC-3926) rescued with a truncated PF6 gene

Goduti DJ & Smith EF (2012) Analyses of functional domains within the PF6 protein of the central apparatus reveal a role for PF6 sub-complex members in regulating flagellar beat frequency. Cytoskeleton 69:179-94

  • Locus:
  • PF6
  • Chromosome:
  • 10

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

pf6-2 (CC-3926) rescued with a truncated PF6 gene

Goduti DJ & Smith EF (2012) Analyses of functional domains within the PF6 protein of the central apparatus reveal a role for PF6 sub-complex members in regulating flagellar beat frequency. Cytoskeleton 69:179-94

  • Locus:
  • PF6
  • Chromosome:
  • 10

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

pf6-2 (CC-3926) rescued with a truncated PF6 gene

Goduti DJ & Smith EF (2012) Analyses of functional domains within the PF6 protein of the central apparatus reveal a role for PF6 sub-complex members in regulating flagellar beat frequency. Cytoskeleton 69:179-94

  • Locus:
  • PF6
  • Chromosome:
  • 10

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

pf6-2 (CC-3926) rescued with a truncated PF6 gene

Goduti DJ & Smith EF (2012) Analyses of functional domains within the PF6 protein of the central apparatus reveal a role for PF6 sub-complex members in regulating flagellar beat frequency. Cytoskeleton 69:179-94

  • Locus:
  • PF6
  • Chromosome:
  • 10

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

FAP74amiRNA mated with pf28

DiPetrillo CG & Smith EF (2011) The Pcdp1 complex coordinates the activity of dynein isoforms to produce wild-type ciliary motility. Mol Biol Cell. 22:4527-38

  • Locus:
  • FAP74, PF28 [DHC15]
  • Chromosome:
  • 6, 11

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

FAP74amiRNA mated with ida1

DiPetrillo CG & Smith EF (2011) The Pcdp1 complex coordinates the activity of dynein isoforms to produce wild-type ciliary motility. Mol Biol Cell. 22:4527-38

  • Locus:
  • FAP74, IDA1 [DHC1]
  • Chromosome:
  • 6, 12

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

FAP74amiRNA mated with ida5

DiPetrillo CG & Smith EF (2011) The Pcdp1 complex coordinates the activity of dynein isoforms to produce wild-type ciliary motility. Mol Biol Cell. 22:4527-38

  • Locus:
  • FAP74, IDA5
  • Chromosome:
  • 6, 13

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of katanin p60

Dymek EE & Smith EF (2012) PF19 encodes the p60 catalytic subunit of katanin and is required for assembly of the flagellar central apparatus in Chlamydomonas. J Cell Sci. 125:3357-66

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

amiRNA knockdown of katanin p60

Dymek EE & Smith EF (2012) PF19 encodes the p60 catalytic subunit of katanin and is required for assembly of the flagellar central apparatus in Chlamydomonas. J Cell Sci. 125:3357-6

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

GFP tagged katanin p60

Dymek EE & Smith EF (2012) PF19 encodes the p60 catalytic subunit of katanin and is required for assembly of the flagellar central apparatus in Chlamydomonas. J Cell Sci. 125:3357-6

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

Double mutant lacking both PACRG and FAP20

Dymek EE, Lin J, Fu G, Porter ME, Nicastro D, Smith EF (2019) PACRG and FAP20 form the inner junction of axonemal doublet microtubules and regulate ciliary motility. Mol Biol Cell. 30:1805-1816

  • Locus:
  • PACRG, FAP20 [BUG22]
  • Chromosome:
  • 2, 7

From Erin Dymek, Elizabeth Smith lab, Dartmouth College, November 2019

D2 (pf16 insertional allele) rescued with a C-terminal BCCP tagged PF16

Fu G, Zhao L, Dymek E, Hou Y, Song K, Phan N, Shang Z, Smith EF, Witman GB, Nicastro D (2019) Structural organization of the C1a-e-c supercomplex within the ciliary central apparatus. J Cell Biol. 218:4236-4251

  • Locus:
  • PF16
  • Chromosome:
  • 9

From Thamali Kariyawasam and Jae-Hyeok Lee, University of British Columbia, December 2019

Kariyawasam T, Joo S, Lee J, Toor D, Gao AF, Noh KC, Lee JH (2019) TALE homeobox heterodimer GSM1/GSP1 is a molecular switch that prevents unwarranted genetic recombination in Chlamydomonas. Plant J. 100:938-953

From Thamali Kariyawasam and Jae-Hyeok Lee, University of British Columbia, December 2019

Kariyawasam T, Joo S, Lee J, Toor D, Gao AF, Noh KC, Lee JH (2019) TALE homeobox heterodimer GSM1/GSP1 is a molecular switch that prevents unwarranted genetic recombination in Chlamydomonas. Plant J. 100:938-953

From Thamali Kariyawasam and Jae-Hyeok Lee, University of British Columbia, December 20199

Kariyawasam T, Joo S, Lee J, Toor D, Gao AF, Noh KC, Lee JH (2019) TALE homeobox heterodimer GSM1/GSP1 is a molecular switch that prevents unwarranted genetic recombination in Chlamydomonas. Plant J. 100:938-953

Lee JH, Lin H, Joo S, Goodenough U (2008) Early sexual origins of homeoprotein heterodimerization and evolution of the plant KNOX/BELL family. Cell. 133:829-40

From Thamali Kariyawasam and Jae-Hyeok Lee, University of British Columbia, December 2019

Kariyawasam T, Joo S, Lee J, Toor D, Gao AF, Noh KC, Lee JH (2019) TALE homeobox heterodimer GSM1/GSP1 is a molecular switch that prevents unwarranted genetic recombination in Chlamydomonas. Plant J. 100:938-953

Lee JH, Lin H, Joo S, Goodenough U (2008) Early sexual origins of homeoprotein heterodimerization and evolution of the plant KNOX/BELL family. Cell. 133:829-40

From Evan Cronmiller and Jae-Hyeok Lee, University of British Columbia. December 2019

This strain has high mating and transformation efficiency. It is the progeny of CC-3491 (cw15 mt-) and CC-85 (nic7 mt+), isolate 1926.

Cronmiller E, Toor D, Shao NC, Kariyawasam T, Wang MH, Lee JH (2019) Cell wall integrity signaling regulates cell wall-related gene expression in Chlamydomonas reinhardtii. Scientific reports, 9(1), 12204. doi:10.1038/s41598-019-48523-4

  • Locus:
  • NIC7 [QSA1]
  • Chromosome:
  • 6

From Nitya Subrahmanian, Patrice P. Hamel lab, The Ohio State University, February 2020

Genotype: mt+; arg7-8 

This strain originally came from Dr. Jean David Rochaix (University of Geneva) and is a cross of CC-124 with an arg7-8 mutant strain.

Barbieri MR, Larosa V, Nouet C, Subrahmanian N, Remacle C, Hamel PP. A forward genetic screen identifies mutants deficient for mitochondrial complex I assembly in Chlamydomonas reinhardtii. Genetics. 2011 Jun;188(2):349-58. doi: 10.1534/genetics.111.128827. Epub 2011 Apr 5. PubMed PMID: 21467570; PubMed Central PMCID: PMC3122308.

Subrahmanian N, Castonguay AD, Fatnes TA, Hamel PP. Chlamydomonas reinhardtii as a plant model system to study mitochondrial complex I dysfunction. Plant Direct. 2020 Feb 3;4(2):e00200. doi: 10.1002/pld3.200. eCollection 2020 Feb. PubMed PMID: 32025618; PubMed Central PMCID: PMC6996877.

Subrahmanian N, Castonguay AD, Remacle C, Hamel PP. Assembly of Mitochondrial Complex I Requires the Low-Complexity Protein AMC1 in Chlamydomonas reinhardtii. Genetics. 2020 Feb 19. pii: genetics.303029.2020. doi: 10.1534/genetics.120.303029. [Epub ahead of print] PubMed PMID: 32075865.

From Nitya Subrahmanian, Patrice P. Hamel lab, The Ohio State University, February 2020

Genotype: mt-; arg7-8 

This strain is originally from Dr. Jean David Rochaix (University of Geneva) and is a cross of CC-124 with an arg7-8 mutant strain.

Barbieri MR, Larosa V, Nouet C, Subrahmanian N, Remacle C, Hamel PP. A forward genetic screen identifies mutants deficient for mitochondrial complex I assembly in Chlamydomonas reinhardtii. Genetics. 2011 Jun;188(2):349-58. doi: 10.1534/genetics.111.128827. Epub 2011 Apr 5. PubMed PMID: 21467570; PubMed Central PMCID: PMC3122308.

Subrahmanian N, Castonguay AD, Fatnes TA, Hamel PP. Chlamydomonas reinhardtii as a plant model system to study mitochondrial complex I dysfunction. Plant Direct. 2020 Feb 3;4(2):e00200. doi: 10.1002/pld3.200. eCollection 2020 Feb. PubMed PMID: 32025618; PubMed Central PMCID: PMC6996877.

Subrahmanian N, Castonguay AD, Remacle C, Hamel PP. Assembly of Mitochondrial Complex I Requires the Low-Complexity Protein AMC1 in Chlamydomonas reinhardtii. Genetics. 2020 Feb 19. pii: genetics.303029.2020. doi: 10.1534/genetics.120.303029. [Epub ahead of print] PubMed PMID: 32075865.

From Nitya Subrahmanian, Patrice P. Hamel lab, The Ohio State University, February 2020

Genotype: mt+; arg7-8; nuob10::APHVIII

This strain is originally from Rosario Barbieri/Patrice P. Hamel (OSU) and was created by mutagenesis of 3A+ (CC-5589) with a cassette containing the APHVIII transgene from pSL18. It is a mitochondrial complex I-deficient strain that harbors an insertional mutation in the NUOB10 gene encoding the NUOB10 / PDSW subunit of mitochondrial complex I. It is paromomycin resistant.

This strain should be maintained in low-medium light conditions (max. 50 µmol.m-2.s-1) and the media should contain arginine. Do not maintain in the dark as it is respiratory-deficient.

Barbieri MR, Larosa V, Nouet C, Subrahmanian N, Remacle C, Hamel PP. A forward genetic screen identifies mutants deficient for mitochondrial complex I assembly in Chlamydomonas reinhardtii. Genetics. 2011 Jun;188(2):349-58. doi: 10.1534/genetics.111.128827. Epub 2011 Apr 5. PubMed PMID: 21467570; PubMed Central PMCID: PMC3122308.

Subrahmanian N, Castonguay AD, Fatnes TA, Hamel PP. Chlamydomonas reinhardtii as a plant model system to study mitochondrial complex I dysfunction. Plant Direct. 2020 Feb 3;4(2):e00200. doi: 10.1002/pld3.200. eCollection 2020 Feb. PubMed PMID: 32025618; PubMed Central PMCID: PMC6996877.

Subrahmanian N, Castonguay AD, Remacle C, Hamel PP. Assembly of Mitochondrial Complex I Requires the Low-Complexity Protein AMC1 in Chlamydomonas reinhardtii. Genetics. 2020 Feb 19. pii: genetics.303029.2020. doi: 10.1534/genetics.120.303029. [Epub ahead of print] PubMed PMID: 32075865.

From Nitya Subrahmanian, Patrice P. Hamel lab, The Ohio State University, February 2020

Genotype: mt+; arg7-8; nuob10::APHVIII; NUOB10; ARG7

This is the amc5 strain complemented with the wild-type NUOB10 gene.

Mutagenesis of amc5 with cosmid 7D10 (Rochaix cosmid library) containing the wild-type NUOB10 gene and selected based on ARG7 marker in the cosmid. It is paromomycin resistant and an arginine prototroph.

This strain should be maintained in low-medium light conditions (max. 50 µmol.m-2.s-1).

Subrahmanian N, Castonguay AD, Fatnes TA, Hamel PP. Chlamydomonas reinhardtii as a plant model system to study mitochondrial complex I dysfunction. Plant Direct. 2020 Feb 3;4(2):e00200. doi: 10.1002/pld3.200. eCollection 2020 Feb. PubMed PMID: 32025618; PubMed Central PMCID: PMC6996877.

Subrahmanian N, Castonguay AD, Remacle C, Hamel PP. Assembly of Mitochondrial Complex I Requires the Low-Complexity Protein AMC1 in Chlamydomonas reinhardtii. Genetics. 2020 Feb 19. pii: genetics.303029.2020. doi: 10.1534/genetics.120.303029. [Epub ahead of print] PubMed PMID: 32075865.