Methods for Modulating the Pathway of NF-κB Using Short Hairpin RNA (ShRNA).

Gene Silencing Genetic Vectors / genetics Lentivirus / genetics NF-kappa B / genetics RNA Interference RNA, Small Interfering / genetics
Gene silencing Lentiviral vectors NF-kB RelA ShRNA Transduction

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2021
Historique:
entrez: 8 7 2021
pubmed: 9 7 2021
medline: 13 1 2022
Statut: ppublish

Résumé

Target gene silencing is a strategy that can be used to turn off pathways or genes which are difficult to turn off pharmacologically, both because of lack of targeting drugs, or because of the risk of wider off-target effects. Here we describe the design and use of short hairpin RNA (ShRNA) and lentiviral vectors as an efficient technique for silencing NF-kappaB (NF-κB) pathway in cultured cells. This method can be used also in hard to transfect primary cell cultures.

Identifiants

DOI: 10.1007/978-1-0716-1669-7_6 PMID: 34236634
pubmed: 34236634
doi: 10.1007/978-1-0716-1669-7_6
doi:

Substances chimiques

NF-kappa B 0
RNA, Small Interfering 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

95-107

Références

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doi: 10.2144/05392RV01
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Filion MC, Phillips NC (1997) Toxicity and immunomodulatory activity of liposomal vectors formulated with cationic lipids toward immune effector cells. Biochim Biophys Acta Biomembr. https://doi.org/10.1016/S0005-2736(97)00126-0
Sliva K, Schnierle BS (2010) Selective gene silencing by viral delivery of short hairpin RNA. Virol J 7:248
doi: 10.1186/1743-422X-7-248
Paddison P, Caudy AA, Bernstein E, Hannon GJ, Conklin DS (2002) Short hairpin RNAs (shRNAs) induce sequence specific silencing in mammalian cells. Genes Dev 16:948–958
doi: 10.1101/gad.981002
Paddison PJ, Caudy AA, Hannon GJ (2002) Stable suppression of gene expression by RNAi in mammalian cells. Proc Natl Acad Sci 99:1443–1448
doi: 10.1073/pnas.032652399
Rubinson DA, Dillon CP, Kwiatkowski AV et al (2003) A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference. Nat Genet 33(3):401–406
doi: 10.1038/ng1117
Mauro C, Leow SC, Anso E et al (2011) NF-κB controls energy homeostasis and metabolic adaptation by upregulating mitochondrial respiration. Nat Cell Biol 13(10):1272–1279
doi: 10.1038/ncb2324
Yang J, Feng S, Yi G et al (2016) Inhibition of RelA expression via RNA interference induces immune tolerance in a rat keratoplasty model. Mol Immunol 73:88–97
doi: 10.1016/j.molimm.2016.03.014
Moore CB, Guthrie EH, Huang MT, Taxman DJ (2010) Short hairpin RNA (shRNA): design, delivery, and assessment of gene knockdown. Methods Mol Biol 629:141–158
pubmed: 20387148 pmcid: 3679364
Paddison P, Cleary M, Silva J et al (2004) Cloning of short hairpin RNAs for gene knockdown in mammalian cells. Nat Methods 1:163–167
doi: 10.1038/nmeth1104-163
Knott SRV, Maceli A, Erard N et al (2014) A computational algorithm to predict shRNA potency. Mol Cell 56(6):796–807
doi: 10.1016/j.molcel.2014.10.025

Auteurs

Marta Moretti (M)

Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.

Barbara Di Francesco (B)

Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.

Mauro Di Vito Nolfi (MDV)

Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, L'Aquila, Italy.

Annapaola Angrisani (A)

Department of Molecular Medicine, Sapienza University of Rome, Rome, Italy.

Enrico De Smaele (E)

Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy. enrico.desmaele@uniroma1.it.

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Classifications MeSH

questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Methods for Modulating the Pathway of NF-κB...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Vecteurs génétiques › Methods for Modulating the Pathway of NF-κB...
questionsmedicales.fr › Virus › Virus à ARN › Retroviridae › Lentivirus › Methods for Modulating the Pathway of NF-κB...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Facteurs de transcription › Facteur de transcription NF-kappa B › Methods for Modulating the Pathway of NF-κB...
questionsmedicales.fr › Phénomènes génétiques › Régulation de l'expression des gènes › Épigenèse génétique › Extinction de l'expression des gènes › Interférence par ARN › Methods for Modulating the Pathway of NF-κB...
questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN non traduit › Petit ARN non traduit › Petit ARN interférent › Methods for Modulating the Pathway of NF-κB...