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Plant Cell. 2019 Sep 27. pii: tpc.00454.2019. doi: 10.1105/tpc.19.00454. [Epub ahead of print]

CRISPR-TSKO: A Technique for Efficient Mutagenesis in Specific Cell Types, Tissues, or Organs in Arabidopsis.

Author information

1
VIB - Ghent University CITY: Ghent Belgium [BE].
2
VIB-UGENT Center of Plant Systems Biology CITY: Ghent POSTAL_CODE: 9052 Belgium [BE].
3
VIB-UGent Center for Plant Systems Biology CITY: Gent Belgium [BE].
4
VIB-Ghent University CITY: Ghent Belgium [BE].
5
VIB-UGENT Center of Plant Systems Biology, 9052 Ghent, Belgium CITY: Ghent Belgium [BE].
6
Ghent University CITY: Ghent Belgium [BE].
7
VIB - Ghent University CITY: GENT POSTAL_CODE: 9052 Belgium [BE].
8
VIB-UGENT Center of Plant Systems Biology, 9052 Ghent, Belgium CITY: Ghent POSTAL_CODE: 9052 Belgium [BE].
9
VIB-UGent Center for Plant Systems Biology CITY: Gent POSTAL_CODE: 9052 Belgium [BE] thomas.jacobs@ugent.vib.be.

Abstract

Detailed functional analyses of many fundamentally-important plant genes via conventional loss-of-function approaches are impeded by severe pleiotropic phenotypes. In particular, mutations in genes that are required for basic cellular functions and/or reproduction often interfere with the generation of homozygous mutant plants, precluding further functional studies. To overcome this limitation, we devised a CRISPR-based tissue-specific knockout system, CRISPR-TSKO, enabling the generation of somatic mutations in particular plant cell types, tissues, and organs. In Arabidopsis, CRISPR-TSKO mutations in essential genes caused well-defined, localized phenotypes in the root cap, stomatal lineage, or entire lateral roots. The underlying modular cloning system allows for efficient selection, identification, and functional analysis of mutant lines directly in the first transgenic generation. The efficacy of CRISPR-TSKO opens new avenues to discover and analyze gene functions in spatial and temporal contexts of plant life while avoiding pleiotropic effects of system-wide loss of gene function.

PMID:
31562216
DOI:
10.1105/tpc.19.00454
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