Strains

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Olga Baidukova, Peter Hegemann lab, Humboldt University-Berlin, July 2023

From Saul Purton, University College London, August 2023

Origin: psbH::aadA knockout in which part of the psbH coding region and 3’ UTR is replaced with the aadA cassette through chloroplast transformation

Culture maintenance: Acetate-requiring, light-sensitive (ΔPSII mutant)

This strain was created to serve as a recipient for ‘marker-free’ chloroplast transformation where selection is based on restoration of phototrophy using the WT psbH gene as the marker. This strain has the same psbH::aadA genotype as strain TN72 (see: CC-5168) but was created using the cell-walled wild-type strain CC-1690 rather that a cell-wall deficient strain carrying the cw15 nuclear lesion. HT72 is therefore suitable for transformation using the microparticle bombardment method. 

Wannathong T, Waterhouse JC, Young RE, Economou CK, Purton S. New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii. Appl Microbiol Biotechnol. 2016 Jun;100(12):5467-77. doi: 10.1007/s00253-016-7354-6. Epub 2016 Feb 18. PMID: 26887319; PMCID: PMC4875957.

Larrea-Alvarez M, Young R, Purton S. A Simple Technology for Generating Marker-Free Chloroplast Transformants of the Green Alga Chlamydomonas reinhardtii. Methods Mol Biol. 2021;2317:293-304. doi: 10.1007/978-1-0716-1472-3_17. PMID: 34028777.

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Background: CC-5325 

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Chaux F, Burlacot A, Mekhalfi M, Auroy P, Blangy S, Richaud P, Peltier G. Flavodiiron Proteins Promote Fast and Transient O2 Photoreduction in Chlamydomonas. Plant Physiol. 2017 Jul;174(3):1825-1836. doi: 10.1104/pp.17.00421. Epub 2017 May 9. PMID: 28487478; PMCID: PMC5490913.

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Background: 137AH

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Tolleter D, Ghysels B, Alric J, Petroutsos D, Tolstygina I, Krawietz D, Happe T, Auroy P, Adriano JM, Beyly A, Cuiné S, Plet J, Reiter IM, Genty B, Cournac L, Hippler M, Peltier G. Control of hydrogen photoproduction by the proton gradient generated by cyclic electron flow in Chlamydomonas reinhardtii. Plant Cell. 2011 Jul;23(7):2619-30. doi: 10.1105/tpc.111.086876. Epub 2011 Jul 15. PMID: 21764992; PMCID: PMC3226202.

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Background: flvB_21 crossed with pgrl1

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Background: flvB_21 crossed with pgrl1

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Adrien Burlacot, Carnegie Institution for Science, August 2023

Background: flvB_21 crossed with pgrl1

Origin: Electroporation with pAR-mtHSP70C-Clover-tPSAD, selected on hygromycin 20ug/mL

Culture maintenance: Low light

Algal CO2 capture is powered by alternative electron pathways of photosynthesis. Gilles Peltier, Carolyne Stoffel, Justin Findinier, Sai Kiran Madireddi, Ousmane Dao, Virginie Epting, Arthur Grossman, Yonghua Li-Beisson, Adrien Burlacot. bioRxiv 2023.08.08.552514; doi: https://doi.org/10.1101/2023.08.08.552514

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Normal phenotype

Background: tua1(int1)

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 1 of TUA2 (Cre04.g216850); a hygromycin-resistant gene cassette was inserted into the region near 3’UTR of TUA1 (Cre03.g190950)

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Lacks motility

Background: tua1(int1)

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 1 of TUA2 (Cre04.g216850); a hygromycin-resistant gene cassette was inserted into the region near 3’UTR of TUA1 (Cre03.g190950); four glutamate residues (E445, E447, E449, and E450) in TUA1 C-terminus region are all substituted to alanine

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Normal phenotype

Background: CC-124

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 2 of TUB1 (Cre12.g542250)

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Normal phenotype

Background: tub1(int2)

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 2 of TUB1 (Cre12.g542250); a hygromycin-resistant gene cassette was inserted into the region near 3’UTR of TUB2 (Cre12.g549550)

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Impaired swimming motility

Background: tub1(int2)

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 2 of TUB1 (Cre12.g542250); a hygromycin-resistant gene cassette was inserted into the region near 3’UTR of TUB2 (Cre12.g549550); atop codon was inserted into TUB2 gene to eliminate four glutamate residues (E439, E440, E441, and E442) in the C-terminus region

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Tomohiro Kubo, University of Yamanashi-Japan, August 2023

Phenotype: Impaired swimming motility with short flagella lacking the central-pair microtubules

Background: tub1(int2)

Origin: Generated by a CRISPR/Cas9 mediated gene editing; a paromomycin-resistant gene cassette was inserted into the intron 2 of TUB1 (Cre12.g542250); a hygromycin-resistant gene cassette was inserted into the region near 3’UTR of TUB2 (Cre12.g549550), a stop codon was inserted into TUB2 gene to eliminate six glutamate residues (E431, E432, E433, E439, E440, E441, and E442) in the C-terminus region

Kubo T, Tani Y, Yanagisawa HA, Kikkawa M, Oda T. α- and β-tubulin C-terminal tails with distinct modifications are crucial for ciliary motility and assembly. J Cell Sci. 2023 Aug 15;136(16):jcs261070. doi: 10.1242/jcs.261070. Epub 2023 Aug 17. PMID: 37519241.

From Gui Zhang, Karl Lechtreck lab, University of Georgia, November 2023

This strain, originally from George Witman and Yuqing Hou, is a kap null mutant generated in g1/CC-5415. 

Lechtreck KF. Methods for Studying Movement of Molecules Within Cilia. Methods Mol Biol. 2016;1454:83-96. doi: 10.1007/978-1-4939-3789-9_6. PMID: 27514917; PMCID: PMC5269603.

Li S, Wan KY, Chen W, Tao H, Liang X, Pan J. Functional exploration of heterotrimeric kinesin-II in IFT and ciliary length control in Chlamydomonas. Elife. 2020 Oct 28;9:e58868. doi: 10.7554/eLife.58868. PMID: 33112235; PMCID: PMC7652414.

  • Locus:
  • FLA3
  • Chromosome:
  • 10

From Gui Zhang, Karl Lechtreck lab, University of Georgia, November 2023

This is kap null mutant was rescued with sfGFP-KAP (Patel et al.) in the pBR-2xsfGFP backbone (described in Lechtreck 2016) and is zeocin resistant.

Lechtreck KF. Methods for Studying Movement of Molecules Within Cilia. Methods Mol Biol. 2016;1454:83-96. doi: 10.1007/978-1-4939-3789-9_6. PMID: 27514917; PMCID: PMC5269603.

Li S, Wan KY, Chen W, Tao H, Liang X, Pan J. Functional exploration of heterotrimeric kinesin-II in IFT and ciliary length control in Chlamydomonas. Elife. 2020 Oct 28;9:e58868. doi: 10.7554/eLife.58868. PMID: 33112235; PMCID: PMC7652414.

  • Locus:
  • FLA3
  • Chromosome:
  • 10

From Gui Zhang, Karl Lechtreck lab, University of Georgia, November 2023

CC-1396 fla8 mt- was rescued with the FLA8-YFP construct described in Li et al. 2020 and is hygromycin resistant. Isolate A4.

Lechtreck KF. Methods for Studying Movement of Molecules Within Cilia. Methods Mol Biol. 2016;1454:83-96. doi: 10.1007/978-1-4939-3789-9_6. PMID: 27514917; PMCID: PMC5269603.

Li S, Wan KY, Chen W, Tao H, Liang X, Pan J. Functional exploration of heterotrimeric kinesin-II in IFT and ciliary length control in Chlamydomonas. Elife. 2020 Oct 28;9:e58868. doi: 10.7554/eLife.58868. PMID: 33112235; PMCID: PMC7652414.

  • Locus:
  • FLA8
  • Chromosome:
  • 12

From Gui Zhang, Karl Lechtreck lab, University of Georgia, November 2023

CC-1396 fla8 mt- was rescued with the FLA8-YFP construct described in Li et al. 2020 and is hygromycin resistant. Isolate A12.

Lechtreck KF. Methods for Studying Movement of Molecules Within Cilia. Methods Mol Biol. 2016;1454:83-96. doi: 10.1007/978-1-4939-3789-9_6. PMID: 27514917; PMCID: PMC5269603.

Li S, Wan KY, Chen W, Tao H, Liang X, Pan J. Functional exploration of heterotrimeric kinesin-II in IFT and ciliary length control in Chlamydomonas. Elife. 2020 Oct 28;9:e58868. doi: 10.7554/eLife.58868. PMID: 33112235; PMCID: PMC7652414.

  • Locus:
  • FLA8
  • Chromosome:
  • 12

from Kenjiro Yoshimura, Shibaura Institute of Technology-Japan, 3/13/24