Osmani Zhila, Jin Shinnosuke, Mikami Masafumi, Endo Masaki, Atarashi Hiroki, Fujino Kaien, Yamada Tetsuya, Nakahara Kenji S
Research Faculty of Agriculture, Hokkaido University, Sapporo, Japan.
Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran.
Methods Mol Biol. 2019;2028:153-165. doi: 10.1007/978-1-4939-9635-3_9.
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.
一种烟草类钙调蛋白，即rgs-CaM，已被证明能以多种方式与病毒相互作用；它有助于双生病毒感染，但也参与对黄瓜花叶病毒的引发免疫。序列相似性搜索揭示了拟南芥和其他茄科植物（包括马铃薯（Solanum tuberosum））中几种与rgs-CaM相似的类钙调蛋白（rCML）。为了分析每个rCML的功能，使用CRISPR/Cas9系统对马铃薯rCML进行了突变。在此，我们描述了在稳定转化的马铃薯植株中进行CRISPR/Cas9介导的靶向诱变的方案。
