[1]李鹏.利用CRISPR/Cas9系统编辑拟南芥ILR3基因及功能验证[J].江苏农业科学,2020,48(14):78-82.
　Li Peng.Editing gene ILR3 in Arabidopsis using CRISPR/Cas9 system and its function verification[J].Jiangsu Agricultural Sciences,2020,48(14):78-82.
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利用CRISPR/Cas9系统编辑拟南芥ILR3基因
及功能验证()

《江苏农业科学》[ISSN:1002-1302/CN:32-1214/S]

卷:
第48卷
期数:
2020年第14期
页码:
78-82
栏目:
生物技术
出版日期:
2020-07-20

文章信息/Info

Title:
Editing gene ILR3 in Arabidopsis using CRISPR/Cas9 system and its function verification
作者:
李鹏
上海辰山植物园/中国科学院上海辰山植物科学研究中心，上海 201602
Author(s):
Li Peng
关键词:
拟南芥; AtILR3; YAO启动子;  CRISPR/Cas9; 基因编辑
Keywords:
-
分类号:
Q784
DOI:
-
文献标志码:
A
摘要:
拟南芥转录因子ILR3(IAA-Leucine Resistant 3)在铁稳态的调节、葡萄糖异硫氰酸盐(glucosinolate，简称GLS)的生物合成和病原体响应方面起到重要作用。为更深入探索该转录因子在植物体内的更多功能，利用YAO基因启动子驱动Cas9在拟南芥中表达，成功获得ILR3基因编辑突变体。测序结果及序列分析结果表明，在ILR3编辑拟南芥中，该基因编码区发生了碱基缺失或插入，导致蛋白ILR3保守结构域丢失。并且，这些基因编辑突变体T2代幼苗与T-DNA插入突变体ilr3-2在缺铁环境下表现出相同的性状，进一步验证ILR3转录因子在植物对铁的吸收及体内平衡的作用，也为深入研究其更多生物学功能奠定了工作基础。
Abstract:
-

参考文献/References:

［1］Wu W，Yang Y，Lei H. Progress in the application of CRISPR：from gene to base editing［J］. Medicinal Research Reviews，2019，39(2)：665-683.
［2］Strecker J，Jones S，Koopal B，et al. Engineering of CRISPR-Cas12b for human genome editing［J］. Nature Communications，2019，10(1)：212.
［3］Li T，Zhu L，Xiao B，et al. CRISPR-Cpf1-mediated genome editing and gene regulation in human cells［J］. Biotechnology Advances，2019，37(1)：21-27.
［4］Das A，Sharma N，Prasad M. CRISPR/Cas9：a novel weapon in the arsenal to combat plant diseases［J］. Frontiers in Plant Science，2018，9：2008.
［5］Chao T，Liu Z，Zhang Y，et al. Precise and rapid validation of candidate gene by allele specific knockout with CRISPR/Cas9 in wild mice［J］. Frontiers in Genetics，2019，10：124.
［6］Hidalgo-Cantabrana C，Goh Y J，Barrangou R. Characterization and repurposing of type Ⅰ and type Ⅱ CRISPR-Cas systems in bacteria［J］. Journal of Molecular Biology，2019，431(1)：21-33.
［7］Li H J，Liu N Y，Shi D Q，et al. YAO is a nucleolar WD40-repeat protein critical for embryogenesis and gametogenesis in Arabidopsis［J］. BMC Plant Biology，2010，10(1)：169.
［8］Yan L，Wei S，Wu Y，et al. High-efficiency genome editing in Arabidopsis using YAO promoter-driven CRISPR/Cas9 system［J］. Molecular Plant，2015，8(12)：1820-1823.
［9］Rampey R A，Woodward A W，Hobbs B N，et al. An Arabidopsis basic helix-loop-helix leucine zipper protein modulates metal homeostasis and auxin conjugate responsiveness［J］. Genetics，2006，174(4)：1841-1857.
［10］Zhang J，Liu B，Li M，et al. The bHLH transcription factor bHLH104 interacts with IAA-LEUCINE RESISTANT3 and modulates iron homeostasis in Arabidopsis［J］. Plant Cell，2015，27(3)：787-805.
［11］Samira R，Li B，Kliebenstein D，et al. The bHLH transcription factor ILR3 modulates multiple stress responses in Arabidopsis［J］. Plant Molecular Biology，2018，97(4/5)：297-309.
［12］Davis A M，Hall A，Millar A J，et al. Protocol：streamlined sub-protocols for floral-dip transformation and selection of transformants in Arabidopsis thaliana［J］. Plant Methods，2009，5：3.
［13］李敏，杨双，阮燕晔，等. 拟南芥T-DNA插入突变体atsuo3的PCR鉴定［J］. 植物生理学通讯，2006，42(1)：91-94.
［14］Berker K I，Ozyurt D，Demirata B，et al. Solid-phase extraction spectrophotometric determination of total antioxidant capacity in antioxidant-poor samples by using the ferric-ferrozine method［J］. Analytical Sciences，2017，33(6)：683-689.
［15］Yu Z，Chen Q，Chen W，et al. Multigene editing via CRISPR/Cas9 guided by a single-sgRNA seed in Arabidopsis［J］. Journal of Integrative Plant Biology，2018，60(5)：376-381.
［16］Toda E，Koiso N，Takebayashi A，et al. An efficient DNA- and selectable-marker-free genome-editing system using zygotes in rice［J］. Nature Plants，2019，5(4)：363-368.
［17］Wang B，Zhu L，Zhao B，et al. Development of a haploid-inducer mediated genome editing system for accelerating maize breeding［J］. Molecular Plant，2019，12(4)：597-602.
［18］Liang Z，Chen K，Li T，et al. Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes［J］. Nature Communications，2017，8：14261.
［19］Li C，Nguyen V，Liu J，et al. Mutagenesis of seed storage protein genes in soybean using CRISPR/Cas9［J］. BMC Research Notes，2019，12(1)：176.
［20］Sun L，Williams J S，Li S，et al. S-locus F-box proteins are solely responsible for S-RNase-based self-incompatibility of Petunia pollen［J］. Plant Cell，2018，30(12)：2959-2972.
［21］李莉梅，欧阳乐军，尹爱国，等. 1种大片段敲除巨桉细胞分裂素氧化酶基因的CRISPR载体构建［J］. 江苏农业科学，2018，46(12)：19-22.
［22］Ye G N，Stone D，Pang S Z，et al. Arabidopsis ovule is the target for Agrobacterium in planta vacuum infiltration transformation［J］. Plant Journal，1999，19(3)：249-257.
［23］Feng Z，Zhang B，Ding W，et al. Efficient genome editing in plants using a CRISPR/Cas system［J］. Cell Research，2013，23(10)：1229-1232.
［24］Ma X，Zhang Q，Zhu Q，et al. A robust CRISPR/Cas9 system for convenient，high-efficiency multiplex genome editing in monocot and dicot plants［J］. Molecular Plant，2015，8(8)：1274-1284.
［25］Long T A，Tsukagoshi H，Busch W，et al. The bHLH transcription factor POPEYE regulates response to iron deficiency in Arabidopsis roots［J］. Plant Cell，2010，22(7)：2219-2236.
［26］Selote D，Samira R，Matthiadis A，et al. Iron-binding E3 ligase mediates iron response in plants by targeting basic helix-loop-helix transcription factors［J］. Plant Physiology，2015，167(1)：273-286.
［27］Hindt M N，Akmakjian G Z，Pivarski K L，et al. BRUTUS and its paralogs，BTS LIKE1 and BTS LIKE2，encode important negative regulators of the iron deficiency response in Arabidopsis thaliana［J］. Metallomics，2017，9(7)：876-890.
［28］Li X，Zhang H，Ai Q，et al. Two bHLH transcription factors，bHLH34 and bHLH104，regulate iron homeostasis in Arabidopsis thaliana［J］. Plant Physiology，2016，170(4)：2478-2493.
［29］Liang G，Zhang H，Li X，et al. bHLH transcription factor bHLH115 regulates iron homeostasis in Arabidopsis thaliana［J］. Journal of Experimental Botany，2017，68(7)：1743-1755.
［30］孙勤富，刘东晓，林俐，等. 甘蓝型油菜和甘蓝CRISPR/Cas9编辑效果的快速检测［J］. 中国油料作物学报，2018，40(6)：737-744.

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备注/Memo

备注/Memo:
收稿日期：2019-08-01
基金项目：上海市青年科技英才扬帆计划(编号：19YF1414800)。
作者简介：李鹏(1983—)，男，安徽濉溪人，硕士，工程师，主要从事植物抗逆性研究。E-mail：penglee98@163.com。
更新日期/Last Update: 2020-07-20