Tailor-made mutations in Arabidopsis using zinc finger nucleases.
Zinc finger nucleases (ZFNs) are proteins engineered to make site-specific double-strand breaks (DSBs) in a DNA sequence of interest. Imprecise repair of the ZFN-induced DSBs by the nonhomologous end-joining (NHEJ) pathway results in a spectrum of mutations, such as nucleotide substitutions, insertions, and deletions. Here we describe a method for targeted mutagenesis in Arabidopsis with ZFNs, which are engineered by context-dependent assembly (CoDA). This ZFN-induced mutagenesis method is an alternative to other currently available gene knockout or knockdown technologies and is useful for reverse genetic studies.
Department of Genetics, Cell Biology & Development and Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.
Yiping
Y
Qi
Colby G
CG
Starker
Feng
F
Zhang
Nicholas J
NJ
Baltes
Daniel F
DF
Voytas
D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004252:DNA Mutational Analysis; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis; D030821:Plants, Genetically Modified / Q000235:genetics*; D014170:Transformation, Genetic; D016335:Zinc Fingers
D018744:DNA, Plant; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific