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CRISPR/Cas9-Mediated Editing of Genes Encoding rgs-CaM-like Proteins in Transgenic Potato Plants.

Methods Mol. Biol.. 2019-01; 
OsmaniZhila,JinShinnosuke,MikamiMasafumi,EndoMasaki,AtarashiHiroki,FujinoKaien,YamadaTetsuya,NakaharaKen
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Codon Optimization … Vector and gene constructs for targeted gene mutation. The Cas9 gene sequence of codon-optimized for tobacco and the sequence of the Arabidopsis U6-26 promoter with gRNA scaffolds was synthesized from GenScript (Nanjing) … Get A Quote

摘要

A tobacco calmodulin-like protein, rgs-CaM, has been shown to interact with viruses in a variety of ways; it contributes to geminivirus infections but is also involved in primed immunity to the cucumber mosaic virus. Sequence similarity searches revealed several calmodulin-like proteins similar to rgs-CaM (rCML) in Arabidopsis and other Solanaceae plants, including potato (Solanum tuberosum). To analyze the functions of each rCML, mutations were introduced into potato rCMLs using the CRISPR/Cas9 system. Here, we describe our protocol of the CRISPR/Cas9-mediated targeted mutagenesis in stably transformed potato plants.

关键词

CRISPR/Cas9,Calmodulin-like protein,Potato plants,Targeted mutagenesis,rgs