利用RNA干扰技术对涡虫进行基因敲低
Gene knockdown in planarians using RNA interference.
作者信息
Oviedo Néstor J, Nicolas Cindy L, Adams Dany S, Levin Michael
机构信息
Center for Regenerative and Developmental Biology, Forsyth Institute and Developmental Biology Department, Harvard School of Dental Medicine, Boston, MA 02115, USA.
出版信息
CSH Protoc. 2008 Oct 1;2008:pdb.prot5054. doi: 10.1101/pdb.prot5054.
Abstract
INTRODUCTIONThis protocol describes how to produce gene knockdown in planarians using RNA interference (RNAi). It is a standard technique to evaluate gene function during regeneration and tissue maintenance in planarians. The procedure involves microinjecting double-stranded RNA (dsRNA) synthesized in vitro. Depending on the gene target, this technique can produce robust phenotypes that can be further evaluated by diverse macroscopic or microscopic procedures.
摘要
引言
本方案描述了如何利用RNA干扰（RNAi）在涡虫中实现基因敲低。这是一种用于评估涡虫再生和组织维持过程中基因功能的标准技术。该程序包括显微注射体外合成的双链RNA（dsRNA）。根据基因靶点的不同，该技术可产生明显的表型，可通过各种宏观或微观程序进一步评估。
相似文献
1
Gene knockdown in planarians using RNA interference.利用RNA干扰技术对涡虫进行基因敲低
CSH Protoc. 2008 Oct 1;2008:pdb.prot5054. doi: 10.1101/pdb.prot5054.
2
RNA interference by feeding in vitro-synthesized double-stranded RNA to planarians: methodology and dynamics.经口饲喂体外合成的双链 RNA 对涡虫的 RNA 干扰：方法学与动力学。
Dev Dyn. 2013 Jun;242(6):718-30. doi: 10.1002/dvdy.23950. Epub 2013 Apr 1.
3
Usefulness of multiple chalk-based food colorings for inducing better gene silencing by feeding RNA interference in planarians.使用多种粉笔状食用色素通过喂食 RNA 干扰在扁形动物中诱导更好的基因沉默的有用性。
Dev Growth Differ. 2018 Jan;60(1):76-81. doi: 10.1111/dgd.12413. Epub 2017 Dec 21.
4
RNA Interference in Planarians: Feeding and Injection of Synthetic dsRNA.涡虫中的RNA干扰：合成双链RNA的喂食与注射
Methods Mol Biol. 2018;1774:455-466. doi: 10.1007/978-1-4939-7802-1_18.
5
RNAi Screening to Assess Tissue Regeneration in Planarians.利用 RNAi 筛选技术评估涡虫组织再生。
Methods Mol Biol. 2022;2450:509-527. doi: 10.1007/978-1-0716-2172-1_27.
6
Systemic RNA Interference in Planarians by Feeding of dsRNA Containing Bacteria.通过喂食含双链RNA的细菌在涡虫中实现系统性RNA干扰
Methods Mol Biol. 2018;1774:445-454. doi: 10.1007/978-1-4939-7802-1_17.
7
Functional analysis of ADARs in planarians supports a bilaterian ancestral role in suppressing double-stranded RNA-response.秀丽隐杆线虫 ADARs 的功能分析支持在抑制双链 RNA 反应中具有两侧对称动物祖先的作用。
PLoS Pathog. 2022 Jan 18;18(1):e1010250. doi: 10.1371/journal.ppat.1010250. eCollection 2022 Jan.
8
A simple "soaking method" for RNA interference in the planarian Dugesia japonica.一种用于日本三角涡虫RNA干扰的简单“浸泡法”。
Dev Genes Evol. 2003 Apr;213(3):138-41. doi: 10.1007/s00427-003-0310-3. Epub 2003 Mar 12.
9
COE loss-of-function analysis reveals a genetic program underlying maintenance and regeneration of the nervous system in planarians.COE功能丧失分析揭示了涡虫神经系统维持和再生背后的遗传程序。
PLoS Genet. 2014 Oct 30;10(10):e1004746. doi: 10.1371/journal.pgen.1004746. eCollection 2014 Oct.
10
Novel monoclonal antibodies to study tissue regeneration in planarians.用于研究涡虫组织再生的新型单克隆抗体。
BMC Dev Biol. 2015 Jan 21;15:2. doi: 10.1186/s12861-014-0050-9.
引用本文的文献
1
Restoration of DNA integrity and the cell cycle by electric stimulation in planarian tissues damaged by ionizing radiation.电刺激在电离辐射损伤的涡虫组织中修复 DNA 完整性和细胞周期。
J Cell Sci. 2022 May 1;135(9). doi: 10.1242/jcs.259304. Epub 2022 May 13.
2
Epithelial Infection With Elicits a Multi-System Response in Planarians.上皮感染在涡虫中引发多系统反应。
Front Microbiol. 2021 Jan 14;11:629526. doi: 10.3389/fmicb.2020.629526. eCollection 2020.
3
Sirtuin-1 regulates organismal growth by altering feeding behavior and intestinal morphology in planarians.Sirtuin-1 通过改变扁形动物的摄食行为和肠道形态来调节生物生长。
J Cell Sci. 2020 May 27;133(10):jcs239467. doi: 10.1242/jcs.239467.
4
Poly(ADP-Ribose) Polymerase-3 Regulates Regeneration in Planarians.多聚（ADP-核糖）聚合酶 3 调控涡虫再生。
Int J Mol Sci. 2020 Jan 29;21(3):875. doi: 10.3390/ijms21030875.
5
Live imaging of intracellular pH in planarians using the ratiometric fluorescent dye SNARF-5F-AM.使用比率荧光染料SNARF-5F-AM对涡虫进行细胞内pH的实时成像。
Biol Methods Protoc. 2019;4(1):bpz005. doi: 10.1093/biomethods/bpz005. Epub 2019 May 29.
6
The Role of Early Bioelectric Signals in the Regeneration of Planarian Anterior/Posterior Polarity.早期生物电信号在水螅前后极性再生中的作用。
Biophys J. 2019 Mar 5;116(5):948-961. doi: 10.1016/j.bpj.2019.01.029. Epub 2019 Feb 1.
7
Cysteine and Aspartyl Proteases Contribute to Protein Digestion in the Gut of Freshwater Planaria.半胱氨酸和天冬氨酸蛋白酶有助于淡水涡虫肠道中的蛋白质消化。
PLoS Negl Trop Dis. 2016 Aug 8;10(8):e0004893. doi: 10.1371/journal.pntd.0004893. eCollection 2016 Aug.
8
The Akt signaling pathway is required for tissue maintenance and regeneration in planarians.Akt信号通路是涡虫组织维持和再生所必需的。
BMC Dev Biol. 2016 Apr 11;16:7. doi: 10.1186/s12861-016-0107-z.
9
Regional signals in the planarian body guide stem cell fate in the presence of genomic instability.在基因组不稳定的情况下，涡虫体内的区域信号引导干细胞命运。
Development. 2016 May 15;143(10):1697-709. doi: 10.1242/dev.131318. Epub 2016 Mar 24.
10
Planarians as a model of aging to study the interaction between stem cells and senescent cells in vivo.涡虫作为衰老模型用于研究体内干细胞与衰老细胞之间的相互作用。
Pathobiol Aging Age Relat Dis. 2015 Dec 10;5:30052. doi: 10.3402/pba.v5.30052. eCollection 2015.
本文引用的文献
1
Live Imaging of Planarian Membrane Potential Using DiBAC4(3).使用DiBAC4(3)对涡虫膜电位进行实时成像。
CSH Protoc. 2008 Oct 1;2008:pdb.prot5055. doi: 10.1101/pdb.prot5055.
2
Establishing and maintaining a colony of planarians.建立并维持涡虫种群。
CSH Protoc. 2008 Oct 1;2008:pdb.prot5053. doi: 10.1101/pdb.prot5053.
3
Planarians: a versatile and powerful model system for molecular studies of regeneration, adult stem cell regulation, aging, and behavior.涡虫：用于再生、成体干细胞调控、衰老及行为分子研究的通用且强大的模型系统。
CSH Protoc. 2008 Oct 1;2008:pdb.emo101. doi: 10.1101/pdb.emo101.
4
Double-stranded RNA specifically disrupts gene expression during planarian regeneration.双链RNA在涡虫再生过程中特异性地破坏基因表达。
Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5049-54. doi: 10.1073/pnas.96.9.5049.
Zotero 插件