利用CRISPR/Cas9基因编辑技术构建水稻ospin9突变体

吴世洋,杨晓祎,张艳雯,侯典云,胥华伟

Generation of ospin9 Mutants in Rice by CRISPR/Cas9 Genome Editing Technology

WU ShiYang,YANG XiaoYi,ZHANG YanWen,HOU DianYun,XU HuaWei

表1 引物及序列

Table 1 The primers and sequence in this study

引物名称 Primer name 引物序列 Primer sequence (5′-3′) 用途 Usage PIN9-CRISPR-F TGTGTTTCTCCAACGAGCAGTGCGC CRISPR/Cas9载体构建 CRISPR/Cas9 vector construction PIN9-CRISPR-R AAACGCGCACTGCTCGTTGGAGAA M13-F GTTGTAAAACGACGGCCAGTGCC 筛选阳性克隆 Screening for positive clones HPT-F CTGAACTCACCGCGACGTCTGTC 筛选阳性植株 Screening for positive transgenic plants HPT-R TAGCGCGTCTGCTGCTCCATACA PIN9-Assay-F CGACCTGGCTTACGAACGAA 扩增OsPIN9基因组片段 Amplification of OsPIN9 genome fragment PIN9-Assay-R CCATGTCGAAGATGAGCACC PIN1a-qF CCTGAAATCCATCTCCATCCTC OsPIN1a表达分析 Expression analysis of OsPIN1a PIN1a-qR AACGTCGCCACCTTGTT PIN1b-qF GAATCGTGCCCTTTGTGTTTG OsPIN1b表达分析 Expression analysis of OsPIN1b PIN1b-qR TGTAGTAGACGAGGGTGATAGG PIN1c-qF GAGCAATCAGCATCCCGAATA OsPIN1c表达分析 Expression analysis of OsPIN1c PIN1c-qR GAGCAATCAGCATCCCGAATA PIN2-qF CGTCTCCTTCAGGTGGAATATC OsPIN2表达分析 Expression analysis of OsPIN2 PIN2-qR AGAGCCATGAACAAGCCTAAG PIN5a-qF CCCTACCTCAATCCATCACATC OsPIN5a表达分析 Expression analysis of OsPIN5a PIN5a-qR GTAGGGAGACAAGCATTCCAA PIN5b-qF GCAAAGGAGTATGGGCTTCA OsPIN5b表达分析 Expression analysis of OsPIN5b PIN5b-qR GCAATCAGAATCGGCAGAGA PIN9-qF GAGGACTCTCTGTTCACCATTC OsPIN9表达分析 Expression analysis of OsPIN9 PIN9-qR GAGAACGACGCTATCTTGTATCC OsACTIN1-qF CTTCATAGGAATGGAAGCTGCG qRT-PCR内参基因 Internal control for qRT-PCR OsACTIN1-qF CACCTTGATCTTCATGCTGCTA