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Cited by 355

The DOI matches its Crossref record; 141 of the 141 references it cites were found in a registry or an index.

An Indexed, Mapped Mutant Library Enables Reverse Genetics Studies of Biological Processes in Chlamydomonas reinhardtii

Open access
Li, Xiaobo, Zhang, Ru, Patena, Weronika, Gang, Spencer S., Blum, Sean R., Ivanova, Nina, Yue, Rebecca, Robertson, Jacob M., Lefebvre, Paul A., Fitz-Gibbon, Sorel T., Grossman, Arthur R., Jonikas, Martin C.·2016·The Plant Cell

Abstract

The green alga Chlamydomonas reinhardtii is a leading unicellular model for dissecting biological processes in photosynthetic eukaryotes. However, its usefulness has been limited by difficulties in obtaining mutants in specific genes of interest. To allow generation of large numbers of mapped mutants, we developed high-throughput methods that (1) enable easy maintenance of tens of thousands of Chlamydomonas strains by propagation on agar media and by cryogenic storage, (2) identify mutagenic insertion sites and physical coordinates in these collections, and (3) validate the insertion sites in pools of mutants by obtaining >500 bp of flanking genomic sequences. We used these approaches to construct a stably maintained library of 1935 mapped mutants, representing disruptions in 1562 genes. We further characterized randomly selected mutants and found that 33 out of 44 insertion sites (75%) could be confirmed by PCR, and 17 out of 23 mutants (74%) contained a single insertion. To demonstrate the power of this library for elucidating biological processes, we analyzed the lipid content of mutants disrupted in genes encoding proteins of the algal lipid droplet proteome. This study revealed a central role of the long-chain acyl-CoA synthetase LCS2 in the production of triacylglycerol from de novo-synthesized fatty acids.

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Metadata

Authors
Li, X., Zhang, R., Patena, W., Gang, S. S., Blum, S. R., Ivanova, N., Yue, R., Robertson, J. M., Lefebvre, P. A., Fitz-Gibbon, S. T., Grossman, A. R., & Jonikas, M. C.
Venue
The Plant Cell
Published
2016-01-13
Volume
28
Issue
2
Pages
367-387
ISSN
  • 1040-4651 (ISSN-L)
  • 1532-298X
OpenAlex
W2236075974
Cited by
355
References
141
FWCI
12.16
Type
journal article
Language
en
Open access
hybrid

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CoChat. (2026). Citation check: An Indexed, Mapped Mutant Library Enables Reverse Genetics Studies of Biological Processes in Chlamydomonas reinhardtii. https://app.cochat.ai/papers/10.1105/tpc.15.00465
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Li, X., Zhang, R., Patena, W., Gang, S. S., Blum, S. R., Ivanova, N., Yue, R., Robertson, J. M., Lefebvre, P. A., Fitz-Gibbon, S. T., Grossman, A. R., & Jonikas, M. C. (2016). An Indexed, Mapped Mutant Library Enables Reverse Genetics Studies of Biological Processes in Chlamydomonas reinhardtii. The Plant Cell. https://doi.org/10.1105/tpc.15.00465

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