Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (5): 141-153.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1452
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GAO Peng-fei1(), XI Fei-hu1,2, ZHANG Ze-yu1, HU Kai-qiang1, CHEN Kai2, WEI Wen-tao1, DING Jia-zhi1, GU Lian-feng1()
Received:
2020-11-27
Online:
2021-05-26
Published:
2021-06-11
Contact:
GU Lian-feng
E-mail:pfgao925@163.com;lfgu@fafu.edu.cn
GAO Peng-fei, XI Fei-hu, ZHANG Ze-yu, HU Kai-qiang, CHEN Kai, WEI Wen-tao, DING Jia-zhi, GU Lian-feng. Research Progress of Plant VIGS Technology and Its Application in Forestry Science[J]. Biotechnology Bulletin, 2021, 37(5): 141-153.
Fig.1 Molecular mechanism of VIGS (1) Construction of viral vectors. (2) Infection into plant cells. (3) Viral vectors are transcribed in the cytoplasm. (4) Synthesis of dsRNA by RDRP. (5)-(7) Dicer cut dsRNA into siRNA and then loaded it in RISC. (8)-(9) Activation of RISC target specific recognition of target genes resulted in host mRNA degradation and loss of gene expression
Fig. 2 Schematic diagram of VIGS infection technology A: The fragment of target gene (e.g. PDS) was cloned into the VIGS vector. B: Agrobacterium transformation culture PTRV1 and PTRV2 were mixed in equal proportion after resuspended. C: The photobleaching phenotype was observed after about 3 weeks after transformation into seedlings by means of toothpick, vacuum osmosis, or syringe injection to trigger plant defense mechanisms
软件名称 Software name | 软件介绍 Software introduction | 参考文献 Reference |
---|---|---|
SGN | 面向用户快速友好的交互式Web工具,在计算机中模拟细胞中发生的VIGS过程。帮助研究人员更快地设计VIGS构建载体,使用SGN工具可帮助选择沉默区域,识别最可能的目标和非目标位置,还可以进行预测沉默效率 | [ |
pssRNAit | 利用全基因组数据设计特异性siRNA,可以进行全基因组脱靶评估是用于设计植物RNAi的有效的网络服务器工具。开发了针对RNAi途径的每个步骤的计算模型,有助设计沉默构建体 | [ |
siRNA-Scan | 具有多个集成组件的搜索环境,包括序列相似性搜索,以预测载体序列是否会潜在脱靶和脱靶效率。可以搜索数据库中的其他脱靶基因然后选择特异性序列,用来设计具有最小脱靶概率的VIGS载体 | [ |
DESIR | 设计siRNA的工具,基于siRNA反义链位置特异性核苷酸序列位置的线性模型进行设计。可得出siRNA设计结果以及预测沉默结果 http://biodev.extra.cea.fr/DSIR/DSIR.html | [ |
DesiRm | 用于设计能够高效沉默基因的siRNA,可以提交mRNA以找出针对其的有效siRNA,以及提交siRNA及其靶序列以通过核苷酸错配设计更有效的siRNA | [ |
i-Score | 对基因沉默RNA折叠剪切后所产生的siRNA进行效率检测,结果由高到低进行排序 | [ |
DEQOR | 对基因区域能够发生较高沉默的区域进行预测,通过所选物种的转录组和基因组进行BLAST搜索,评估输入的靶基因位置的沉默效率 | [ |
P-SAMS | web应用程序设计人工microRNA,有助于特异性的靶向不同的双子叶和单子叶植物物种中的一个以及多个基因 | [ |
WMD3 | 基于web应用程序可用于设计人工microRNA构建基因沉默表达载体,可用目标搜索找到潜在的靶基因 | [ |
PhieCBEs | 新型,高效,广靶向的植物编辑系统,是水稻和其他植物(作物)的单碱基编辑有效实用的工具,可在基因功能筛选,大规模饱和突变,编辑调控元件,RNA可变剪接,基因沉默等研究中广泛应用 | [ |
Table 1 Gene silence related design software
软件名称 Software name | 软件介绍 Software introduction | 参考文献 Reference |
---|---|---|
SGN | 面向用户快速友好的交互式Web工具,在计算机中模拟细胞中发生的VIGS过程。帮助研究人员更快地设计VIGS构建载体,使用SGN工具可帮助选择沉默区域,识别最可能的目标和非目标位置,还可以进行预测沉默效率 | [ |
pssRNAit | 利用全基因组数据设计特异性siRNA,可以进行全基因组脱靶评估是用于设计植物RNAi的有效的网络服务器工具。开发了针对RNAi途径的每个步骤的计算模型,有助设计沉默构建体 | [ |
siRNA-Scan | 具有多个集成组件的搜索环境,包括序列相似性搜索,以预测载体序列是否会潜在脱靶和脱靶效率。可以搜索数据库中的其他脱靶基因然后选择特异性序列,用来设计具有最小脱靶概率的VIGS载体 | [ |
DESIR | 设计siRNA的工具,基于siRNA反义链位置特异性核苷酸序列位置的线性模型进行设计。可得出siRNA设计结果以及预测沉默结果 http://biodev.extra.cea.fr/DSIR/DSIR.html | [ |
DesiRm | 用于设计能够高效沉默基因的siRNA,可以提交mRNA以找出针对其的有效siRNA,以及提交siRNA及其靶序列以通过核苷酸错配设计更有效的siRNA | [ |
i-Score | 对基因沉默RNA折叠剪切后所产生的siRNA进行效率检测,结果由高到低进行排序 | [ |
DEQOR | 对基因区域能够发生较高沉默的区域进行预测,通过所选物种的转录组和基因组进行BLAST搜索,评估输入的靶基因位置的沉默效率 | [ |
P-SAMS | web应用程序设计人工microRNA,有助于特异性的靶向不同的双子叶和单子叶植物物种中的一个以及多个基因 | [ |
WMD3 | 基于web应用程序可用于设计人工microRNA构建基因沉默表达载体,可用目标搜索找到潜在的靶基因 | [ |
PhieCBEs | 新型,高效,广靶向的植物编辑系统,是水稻和其他植物(作物)的单碱基编辑有效实用的工具,可在基因功能筛选,大规模饱和突变,编辑调控元件,RNA可变剪接,基因沉默等研究中广泛应用 | [ |
树种Species | 病毒载体来源 Viral vector source | 参考文献 Reference |
---|---|---|
胡杨(Populus euphratica) | TRV | [ |
银灰杨(Populus canescens) | TRV | [ |
麻风树(Jatropha curcas) | TRV | [ |
苹果(Malus domestica) | ALSV | [ |
木薯(Manihot esculenta) | ACMV | [ |
梨(Pyrus betulaefolia) 桃(Prunus persica) | ALSV,TRV TRV | [ [ |
樱桃(Prunus avium) | TRV | [ |
荔枝(Litchi chinensis) | TRV | [ |
欧洲越橘(Vaccinium myrtillus) | TRV | [ |
凤丹(Paeonia ostii) | TRV | [ |
橄榄(Oleaceae hoffmanns) | TRV | [ |
喜树(Camptotheca acuminata) | TRV | [ |
紫薇(Lagerstroemia indica) | TRV | [ |
Table 2 Research and application of VIGS system in poplar and other forest tree species
树种Species | 病毒载体来源 Viral vector source | 参考文献 Reference |
---|---|---|
胡杨(Populus euphratica) | TRV | [ |
银灰杨(Populus canescens) | TRV | [ |
麻风树(Jatropha curcas) | TRV | [ |
苹果(Malus domestica) | ALSV | [ |
木薯(Manihot esculenta) | ACMV | [ |
梨(Pyrus betulaefolia) 桃(Prunus persica) | ALSV,TRV TRV | [ [ |
樱桃(Prunus avium) | TRV | [ |
荔枝(Litchi chinensis) | TRV | [ |
欧洲越橘(Vaccinium myrtillus) | TRV | [ |
凤丹(Paeonia ostii) | TRV | [ |
橄榄(Oleaceae hoffmanns) | TRV | [ |
喜树(Camptotheca acuminata) | TRV | [ |
紫薇(Lagerstroemia indica) | TRV | [ |
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