Chinese Journal OF Rice Science ›› 2018, Vol. 32 ›› Issue (3): 219-225.DOI: 10.16819/j.1001-7216.2018.7106
• Research Papers • Previous Articles Next Articles
Xuejiao HU1,2, Jia YANG1,3, Can CHENG1, Jihua ZHOU1, Fuan NIU1, Xinqi WANG1, Meiliang ZHANG2, Liming CAO1,*(), Huangwei CHU1,*()
Received:
2017-09-01
Revised:
2017-12-27
Online:
2018-05-10
Published:
2018-05-10
Contact:
Liming CAO, Huangwei CHU
胡雪娇1,2, 杨佳1,3, 程灿1, 周继华1, 牛付安1, 王新其1, 张美良2, 曹黎明1,*(), 储黄伟1,*()
通讯作者:
曹黎明,储黄伟
基金资助:
CLC Number:
Xuejiao HU, Jia YANG, Can CHENG, Jihua ZHOU, Fuan NIU, Xinqi WANG, Meiliang ZHANG, Liming CAO, Huangwei CHU. Targeted Editing of Rice SD1 Gene UsingCRISPR/Cas9 System[J]. Chinese Journal OF Rice Science, 2018, 32(3): 219-225.
胡雪娇, 杨佳, 程灿, 周继华, 牛付安, 王新其, 张美良, 曹黎明, 储黄伟. 利用CRISPR/Cas9系统定向编辑水稻SD1基因[J]. 中国水稻科学, 2018, 32(3): 219-225.
Add to citation manager EndNote|Ris|BibTeX
URL: http://www.ricesci.cn/EN/10.16819/j.1001-7216.2018.7106
引物名称 Primer name | 引物序列(5′-3′) Primer sequence (5′-3′) |
Oligo-F | TGGCGGAGGATGGAGCCCAAGATCC |
Oligo-R | AAACGGATCTTGGGCTCCATCCTC |
M13-F SEQ1-F SEQ1-R EXON1-F EXON1-R EXON3-F EXON3-R HYG-F HYG-R CAS9-F CAS9-R | TGTAAAACGACGGCCAGT GGGTCATTGATTCGACCATC GTGCTCGGACACCTGGAAGAAC GTCATTGATTCGACCATCATGTCTGTC GAATTACTTGTTCTGTTGCTTCGAAGCA CTCTCCGTTGATGAATGATGATG CTTCTGTTCGTTCCGTTTCGTTC CGTGCTTTCAGCTTCGATGTAGGA GCATATGAAATCACGCCATGTAGTGT CTTCATCGAGCGGATGACCAACT AGATCGTGGTATGTGCCCAGGGA |
Table 1 Primers used in this research.
引物名称 Primer name | 引物序列(5′-3′) Primer sequence (5′-3′) |
Oligo-F | TGGCGGAGGATGGAGCCCAAGATCC |
Oligo-R | AAACGGATCTTGGGCTCCATCCTC |
M13-F SEQ1-F SEQ1-R EXON1-F EXON1-R EXON3-F EXON3-R HYG-F HYG-R CAS9-F CAS9-R | TGTAAAACGACGGCCAGT GGGTCATTGATTCGACCATC GTGCTCGGACACCTGGAAGAAC GTCATTGATTCGACCATCATGTCTGTC GAATTACTTGTTCTGTTGCTTCGAAGCA CTCTCCGTTGATGAATGATGATG CTTCTGTTCGTTCCGTTTCGTTC CGTGCTTTCAGCTTCGATGTAGGA GCATATGAAATCACGCCATGTAGTGT CTTCATCGAGCGGATGACCAACT AGATCGTGGTATGTGCCCAGGGA |
靶位点编号 Targetnumber | 靶位点序列 Sequence of target site | 候选识别位点的毗邻基序Protospacer adjacent motif | 位置 Site / bp | DNA链 DNA strand |
1 | GAGGATGGAGCCCAAGATCC | CGG | 108 | + |
2 | AGATCCCGGAGCCATTCGTG | TGG | 122 | + |
3 | GAGCCATTCGTGTGGCCGAA | CGG | 130 | + |
4 | GGATCTTGGGCTCCATCCTC | AGG | 107 | – |
5 | ACGAATGGCTCCGGGATCTT | GGG | 120 | – |
6 | CACGAATGGCTCCGGGATCT | TGG | 121 | – |
7 | TTCGGCCACACGAATGGCTC | CGG | 129 | – |
Table 2 Putative CRISPR/Cas9 target sites in rice SD1 gene.
靶位点编号 Targetnumber | 靶位点序列 Sequence of target site | 候选识别位点的毗邻基序Protospacer adjacent motif | 位置 Site / bp | DNA链 DNA strand |
1 | GAGGATGGAGCCCAAGATCC | CGG | 108 | + |
2 | AGATCCCGGAGCCATTCGTG | TGG | 122 | + |
3 | GAGCCATTCGTGTGGCCGAA | CGG | 130 | + |
4 | GGATCTTGGGCTCCATCCTC | AGG | 107 | – |
5 | ACGAATGGCTCCGGGATCTT | GGG | 120 | – |
6 | CACGAATGGCTCCGGGATCT | TGG | 121 | – |
7 | TTCGGCCACACGAATGGCTC | CGG | 129 | – |
Fig. 3. Mutation type in SD1 gene of T0transgenic lines. The grey letters indicated the target sequence. The underline indicated protospacer adjacent motif(PAM) sequence. The box represented the insert base. The transverse line indicated deletion sequence. Arrow indicated the Cas9 cleavage site.
Fig. 4. PCR screening of transgenic-free plants in T1 generation. M, D2000marker, W, Wild type;Lanes 1-24, T1 transgenic plants; P1, Primer M13F and Oligo-R;P2, Primer HYG-F and HYG-R; P3, Primer Cas9-F and Cas9-R.
[1] | Doebley J F, Gaut B S, Smith B D.The molecular genetics of crop domestication.Cell, 2006, 127(7): 1309-1321. |
[2] | Liu Q, Xu X, Ren X, Fu H, Wu D, Shu Q.Generation and characterization of low phytic acid germplasm in rice ( Oryza sativa L.).Theor Appl Genet, 2007, 114(5): 803-814. |
[3] | Cho S W, Kim S, Kim J M, Kim J S.Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease.Nat Biotechnol, 2013, 31(3): 230-232. |
[4] | Cong L, Ran F A, Cox D, Lin S, Barretto R, Habib N, Hsu P D, Wu X, Jiang W, Marraffini L A, Zhang F.Multiplex genome engineering using CRISPR/Cas systems. Science, 2013, 339(6121): 819-23. |
[5] | Jinek M, East A, Cheng A, Lin S, Ma E, Doudna J.RNA-programmed genome editing in human cells.Elife, 2013, 2: e00471. |
[6] | Mali P, Aach J, Stranges P B, Esvelt K M, Moosburner M, Kosuri S, Yang L, Church G M.CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering.Nat Biotechnol, 2013, 31(9): 833-838. |
[7] | Mani M, Kandavelou K, Dy F J, Durai S, Chandrasegaran S.Design, engineering, and characterization of zinc finger nucleases.Biochem Biophys Res Commun, 2005, 335(2): 447-457. |
[8] | Bogdanove A J, Voytas D F.TAL effectors: Customizable proteins for DNA targeting.Science, 2011, 333(6051): 1843-1846. |
[9] | Samanta M K, Dey A, Gayen S.CRISPR/Cas9: An advanced tool for editing plant genomes.Transgenic Res, 2016, 25(5): 561-573. |
[10] | Mao Y, Zhang H, Xu N, Zhang B, Gou F, Zhu J K.Application of the CRISPR-Cas system for efficient genome engineering in plants.Mol Plant, 2013, 6(6): 2008-2011. |
[11] | Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi J J, Qiu J L, Gao C.Targeted genome modification of crop plants using a CRISPR-Cas system.Nat Biotechnol, 2013, 31(8): 686-688. |
[12] | Jiang W Z, Zhou H B, Bi H H, Fromm M, Yang B, Weeks D P.Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice.Nucleic Acids Res, 2013, 41(20): e188. |
[13] | Liang Z, Zhang K, Chen K, Gao C.Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system.J Genet Genom, 2014, 41(2): 63-68. |
[14] | Wang Y, Cheng X, Shan Q, Zhang Y, Liu J, Gao C, Qiu J L.Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew.Nat Biotechnol, 2014, 32(9): 947-951. |
[15] | Jacobs T B, LaFayette P R, Schmitz R J, Parrott W A. Targeted genome modifications in soybean with CRISPR/Cas9.BMC Biotechnol, 2015, 15: 16. |
[16] | Li Z, Liu Z B, Xing A, Moon B P, Koellhoffer J P, Huang L, Ward R T, Clifton E, Falco S C, Cigan A M.Cas9-Guide RNA directed genome editing in soybean.Plant Physiol, 2015, 169(2): 960-970. |
[17] | Sun X, Hu Z, Chen R, Jiang Q, Song G, Zhang H, Xi Y.Targeted mutagenesis in soybean using the CRISPR-Cas9 system.Sci Rep, 2015, 5: 10342. |
[18] | Brooks C, Nekrasov V, Lippman Z B, Van Eck J.Efficient gene editing in tomato in the first generation using the clustered regularly interspaced short palindromic repeats/CRISPR-associated9 system.Plant Physiol, 2014, 166(3): 1292-1297. |
[19] | Zhang H, Zhang J, Wei P, Zhang B, Gou F, Feng Z, Mao Y, Yang L, Zhang H, Xu N, Zhu J K.The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation.Plant Biotechnol J, 2014, 12(6): 797-807. |
[20] | Monna L, Kitazawa N, Yoshino R, Suzuki J, Masuda H, Maehara Y, Tanji M, Sato M, Nasu S, Minobe Y.Positional cloning of rice semidwarfing gene,sd-1: Rice “Green Revolution Gene” encodes a mutant enzyme involved in gibberellin synthesis. DNA Res, 2002, 9(1): 11-17. |
[21] | Spielmeyer W, Ellis M H, Chandler P M.Semidwarf (sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene. Proc Natl Acad Sci USA, 2002, 99(13): 9043-9048. |
[22] | Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, Kitano H, Matsuoka M.Green revolution: A mutant gibberellin-synthesis gene in rice.Nature, 2002, 416(6882): 701-702. |
[23] | 谷福林, 翟虎渠, 万建民, 张红生. 水稻矮秆性状研究及矮源育种利用. 江苏农业学报, 2003, 19(1): 48-54. |
Gu F L, Zhai H Q, Wan L M, Zhang H S.Study on Inheritance of dwarf character and its utilization in rice(Oryza sativa L.) breeding. Jiangsu JAgric Sci, 2003, 19(1):48-54. (in Chinese with English abstract) | |
[24] | Asano K, Yamasaki M, Takuno S, Miura K, Katagiri S, Ito T, Doi K, Wu J, Ebana K, Matsumoto T, Innan H, Kitano H, Ashikari M, Matsuoka M. Artificial selection for a green revolution gene during japonica rice domestication.Proc Natl Acad Sci USA#, 2011, 108(27): 11034-11039. |
[25] | Yan M, Zhou S R, Xue H W.CRISPR Primer Designer: Design primers for knockout and chromosome imaging CRISPR-Cas system.J Integr Plant Biol, 2015, 57(7): 613-617. |
[26] | Qiao F, Zhao K J.The Influence of RNAi targeting of OsGA20ox2gene on plant height in rice. Plant Mol Biol Rep, 2011, 29(4): 952. |
[27] | Harrison M M, Jenkins B V, O’Connor-Giles K M, Wildonger J. A CRISPR view of development.Genes Dev, 2014, 28(17): 1859-1872. |
[1] | REN Zhiqi, XUE Kexin, DONG Zheng, LI Xiaoxiang, LI Yongzhao, GUO Yujing, LIU Wenqiang, GUO Liang, SHENG Xinnian, LIU Zhixi, PAN Xiaowu. Identification and Gene Mapping of Outcurved Leaf Mutant ocl1 in Rice [J]. Chinese Journal OF Rice Science, 2023, 37(4): 337-346. |
[2] | XIAO Lequan, LI Lei, DAI Weimin, QIANG Sheng, SONG Xiaoling. Seedling Growth Characteristics of Hybrids Between Transgenic Rice with cry2A*/bar Genes and Weedy Rice [J]. Chinese Journal OF Rice Science, 2023, 37(4): 347-358. |
[3] | LI Gang, GAO Qingsong, LI Wei, ZHANG Wenxia, WANG Jian, CHEN Baoshan, WANG Di, GAO Hao, XU Weijun, CHEN Hongqi, JI Jianhui. Directed Knockout of SD1 Gene Improves Lodging Resistance and Blast Resistance of Rice [J]. Chinese Journal OF Rice Science, 2023, 37(4): 359-367. |
[4] | WANG Shengyong, CHEN Yuhang, CHEN Huili, HUANG Yujie, ZHANG Xiaotian, DING Shuangcheng, WANG Hongwei. Effects of High Temperature on Phenylpropane Metabolism and Downstream Branch Metabolic Pathways in Rice Meiosis [J]. Chinese Journal OF Rice Science, 2023, 37(4): 368-378. |
[5] | HUANG Yaru, XU Peng, WANG Lele, HE Yizhe, WANG Hui, KE Jian, HE Haibing, WU Liquan, YOU Cuicui. Effects of Exogenous Trehalose on Grain Filling Characteristics and Yield Formation of japonica Rice Cultivar W1844 [J]. Chinese Journal OF Rice Science, 2023, 37(4): 379-391. |
[6] | DONG Liqiang, YANG Tiexin, LI Rui, SHANG Wenqi, MA Liang, LI Yuedong, SUI Guomin. Effect of Plant-row Spacing on Rice Yield and Root Morphological and Physiological Characteristics in Super High Yield Field [J]. Chinese Journal OF Rice Science, 2023, 37(4): 392-404. |
[7] | GAO Qianqing, REN Xiaojian, ZHAI Zhongbing, ZHENG Pubing, WU Yuanfen, CUI Kehui. Effects of Panicle and Bud-promoting Nitrogen Fertilizer Application on Growth of Regenerated Bud and Grain Yield of Ratoon Rice [J]. Chinese Journal OF Rice Science, 2023, 37(4): 405-414. |
[8] | HUANG Jinwen, LI Rikun, CHEN Zhicheng, ZHANG Bianhong, LEI Han, PAN Ruixin, YANG Mingyu, PAN Meiqing, TANG Lina. Effects of Straw Returning Techniques on Soil Nutrients, Organic Carbon and Microbial Diversity in Tobacco-rice Rotation System [J]. Chinese Journal OF Rice Science, 2023, 37(4): 415-426. |
[9] | HAN Cong, HE Yuchang, WU Lijuan, JIA Lili, WANG Lei, E Zhiguo. Research Progress in the Function of Basic Leucine Zipper (bZIP) Protein Family in Rice [J]. Chinese Journal OF Rice Science, 2023, 37(4): 436-448. |
[10] | SHEN Yumin, CHEN Mingliang, XIONG Huanjin, XIONG Wentao, WU Xiaoyan, XIAO Yeqing. Phenotypic Analysis and Fine Mapping of blg1(beak like grain 1), a Rice Mutant with Abnormal Palea and Lemma Development [J]. Chinese Journal OF Rice Science, 2023, 37(3): 225-232. |
[11] | DUAN Min, XIE Liujie, GAO Xiuying, TANG Haijuan, HUANG Shanjun, PAN Xiaobiao. Creation of Thermo-sensitive Genic Male Sterile Rice Lines with Wide Compatibility Based on CRISPR/Cas9 Technology [J]. Chinese Journal OF Rice Science, 2023, 37(3): 233-243. |
[12] | WANG Wenting, MA Jiaying, LI Guangyan, FU Weimeng, LI Hubo, LIN Jie, CHEN Tingting, FENG Baohua, TAO Longxing, FU Guanfu, QIN Yebo. Effect of Different Fertilizer Application Rates on Rice Yield and Quality Formation and Its Relationship with Energy Metabolism at High Temperature [J]. Chinese Journal OF Rice Science, 2023, 37(3): 253-264. |
[13] | LIU Aihua, LI Xiaokun. Meta-analysis of Relationship Between Fertilizer Application and Rice Quality [J]. Chinese Journal OF Rice Science, 2023, 37(3): 276-284. |
[14] | YANG Xiaolong, WANG Biao, WANG Benfu, ZHANG Zhisheng, ZHANG Zuolin, YANG Lantian, CHENG Jianping, LI Yang. Effects of Different Water Management on Yield and Rice Quality of Dry-seeded Rice [J]. Chinese Journal OF Rice Science, 2023, 37(3): 285-294. |
[15] | WEI Xiaodong, SONG Xuemei, ZHAO Ling, ZHAO Qingyong, CHEN Tao, LU Kai, ZHU Zhen, HUANG Shengdong, WANG Cailin, ZHANG Yadong. Effects of Silicon and Zinc Fertilizer and Their Application Ways on Yield and Grain Quality of Rice Variety Nanjing 46 [J]. Chinese Journal OF Rice Science, 2023, 37(3): 295-306. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||