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
相似文献
1
Gene knockdown in planarians using RNA interference.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
J Cell Sci. 2020 May 27;133(10):jcs239467. doi: 10.1242/jcs.239467.
4
Poly(ADP-Ribose) Polymerase-3 Regulates Regeneration in Planarians.
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.
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.
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.
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.
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.
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.
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).
CSH Protoc. 2008 Oct 1;2008:pdb.prot5055. doi: 10.1101/pdb.prot5055.
2
Establishing and maintaining a colony of planarians.
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.
CSH Protoc. 2008 Oct 1;2008:pdb.emo101. doi: 10.1101/pdb.emo101.
4
Double-stranded RNA specifically disrupts gene expression during planarian regeneration.
Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):5049-54. doi: 10.1073/pnas.96.9.5049.
Zotero 插件