Name: detection of rna-binding proteins by in vitro rna pull-down in adipocyte culture
Uploaded: 2016-07-22T15:00:00
Description: Watch this Scientific Journal Video about Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture at JoVE.com

Dette arbejde blev finansieret af Singapore NRF fællesskab (NRF-2011NRF-NRFF001-025) samt CBRG (NMRC / CBRG / 0070/2014).

BEMÆRK: RNA&#39;et af interesse i forbindelse med denne undersøgelse er et fragment af AR 3&#39;UTR. 1. Fremstilling af biotin-mærket RNA <ol><li> For at opnå den RNA, forstærke T7-AR-oligo ved PCR, efterfulgt af in vitro transkription ved anvendelse af et kommercielt kit og følge producentens instruktioner 11. BEMÆRK: Primere til PCR er anført i tabel 4: T7-AR-F og T7-AR-R. </li><li> For ikke-specifik binding kontrol, amplificere en dels...

<ol>
	<li>Huot, M. &. #. 2. 0. 1. ;., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sam68+STAR+RNA-binding+protein+regulates+mTOR+alternative+splicing+during+adipogenesis.">The Sam68 STAR RNA-binding protein regulates mTOR alternative splicing during adipogenesis.</a> Mol Cell. 46 (2), 187-199 (2012).</li><li>Sun, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+noncoding+RNAs+regulate+adipogenesis.">Long noncoding RNAs regulate adipogenesis.</a> Proc Natl Acad Sci U S A. 110 (9), 3387-3392 (2013).</li><li>Engreitz, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Xist+lncRNA+exploits+three-dimensional+genome+architecture+to+spread+across+the+X-chromosome.">The Xist lncRNA exploits three-dimensional genome architecture to spread across the X-chromosome.</a> Science. 341 (6147), 1237973 (2013).</li><li>Zhao, X. Y., Li, S., Wang, G. X., Yu, Q., Lin, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+long+noncoding+RNA+transcriptional+regulatory+circuit+drives+thermogenic+adipocyte+differentiation.">A long noncoding RNA transcriptional regulatory circuit drives thermogenic adipocyte differentiation.</a> Mol Cell. 55, 372-382 (2014).</li><li>Alvarez-Dominguez, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=De+novo+reconstruction+of+adipose+tissue+transcriptomes+reveals+long+non-coding+RNA+regulators+of+brown+adipocyte+development.">De novo reconstruction of adipose tissue transcriptomes reveals long non-coding RNA regulators of brown adipocyte development.</a> Cell Metab. 21, 764-776 (2015).</li><li>Adams, D. J., Beveridge, D. J., van der Weyden, L., Mangs, H., Leedman, P. J., Morris, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HADHB,+HuR,+and+CP1+bind+to+the+distal+3-untranslated+region+of+human+renin+mRNA+and+differentially+modulate+renin+expression.">HADHB, HuR, and CP1 bind to the distal 3-untranslated region of human renin mRNA and differentially modulate renin expression.</a> J Biol Chem. 278 (45), 44894-44903 (2003).</li><li>Yeap, B. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+binding+of+HuR+and+poly(C)-binding+protein+to+a+conserved+UC-rich+motif+within+the+3&#180;+untranslated+region+of+the+androgen+receptor+messenger+RNA.">Novel binding of HuR and poly(C)-binding protein to a conserved UC-rich motif within the 3&#180; untranslated region of the androgen receptor messenger RNA.</a> J Biol Chem. 277, 27183-27192 (2002).</li><li>K&#246;nig, J., Zarnack, K., Luscombe, N. M., Ule, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein-RNA+interactions:+new+genomic+technologies+and+perspectives.">Protein-RNA interactions: new genomic technologies and perspectives.</a> Nat Rev Genet. 13, 77-83 (2012).</li><li>Ferr&#232;, F., Colantoni, A., Helmer-Citterich, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revealing+protein-lncRNA+interaction.">Revealing protein-lncRNA interaction.</a> Briefings in Bioinformatics. , 1-11 (2015).</li><li>McHugh, C. A., Russell, P., Guttman, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+comprehensive+experimental+identification+of+RNA-protein+interactions.">Methods for comprehensive experimental identification of RNA-protein interactions.</a> Genome Biol. 15 (1), 203 (2014).</li><li>Lee, Y. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AU-rich+element-binding+protein+negatively+regulates+CCAAT+enhancer+binding+protein+mRNA+stability+during+long+term+synaptic+plasticity+in+Aplysia.">AU-rich element-binding protein negatively regulates CCAAT enhancer binding protein mRNA stability during long term synaptic plasticity in Aplysia.</a> Proc Natl Acad Sci U S A. 109 (38), 15520-15525 (2012).</li><li>Hermann, M., Cermak, T., Voytas, D. F., Pelczar, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+genome+engineering+using+designer+nucleases.">Mouse genome engineering using designer nucleases.</a> J Vis Exp. (86), (2014).</li><li>Tsai, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long+Noncoding+RNA+as+Modular+Scaffold+of+Histone+Modification+Complexes.">Long Noncoding RNA as Modular Scaffold of Histone Modification Complexes.</a> Science. 329 (5992), 689-693 (2010).</li><li>Wan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Landscape+and+variation+of+RNA+secondary+structure+across+the+human+transcriptome.">Landscape and variation of RNA secondary structure across the human transcriptome.</a> Nature. 505, 706-709 (2014).</li><li>Li, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+mRNA+binding+proteins+that+regulate+the+stability+of+LDL+receptor+mRNA+through+AU+rich+elements.">Identification of mRNA binding proteins that regulate the stability of LDL receptor mRNA through AU rich elements.</a> J Lipid Res. 50 (5), 820-831 (2009).</li><li>McHugh, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Xist+lncRNA+interacts+directly+with+SHARP+to+silence+transcription+through+HDAC3.">The Xist lncRNA interacts directly with SHARP to silence transcription through HDAC3.</a> Nature. 521 (7551), 232-236 (2015).</li><li>Chu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systematic+Discovery+of+Xist+RNA+Binding+Proteins.">Systematic Discovery of Xist RNA Binding Proteins.</a> Cell. 161 (2), 404-416 (2015).</li><li>Chu, C., Spitale, R. C., Chang, H. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technologies+to+probe+functions+and+mechanisms+of+long+noncoding+RNAs.">Technologies to probe functions and mechanisms of long noncoding RNAs.</a> Nat Struct Mol Biol. 22 (1), 29-35 (2015).</li><li>Zhao, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+identification+of+Polycomb-associated+RNAs+by+RIP-seq.">Genome-wide identification of Polycomb-associated RNAs by RIP-seq.</a> Mol Cell. 40 (6), 939-953 (2010).</li></ol>

lncRNAs og RBP&#39;er har afgørende roller i sundhed og sygdom, men de molekylære mekanismer i disse molekyler er dårligt forstået. Identifikation af proteiner, der interagerer med lncRNA molekyler er et vigtigt skridt i retning af at belyse de regulatoriske mekanismer. Berigelse af RBP&#39;er af RNA pull-down assay er baseret på in-opløsning indfangning af interagerende kompleks, således at proteiner fra et cellelysat selektivt kan ekstraheres under anvendelse af biotinylerede RNA lokkemad bundet til streptavidi...

A correction to the author list was made to: <a href="http://www.jove.com/video/54207/detection-rna-binding-proteins-vitro-rna-pull-down-adipocyte">Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture</a>.
The author list has been updated from:
Qianfan Bai1*, Zhiqiang Bai1*, Lei Sun1,2
1Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School
2Institute of Molecular and Cell Biology, Singapore
*These authors contributed equally
to:
Qianfan Bai1*, Zhiqiang Bai1*, Shaohai Xu3, Lei Sun1,2
1Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School
2Institute of Molecular and Cell Biology, Singapore
3Division of Bioengineering, School of Chemical &#38; Biomedical Engineering, Nanyang Technological University
*These authors contributed equally

A correction to the author list was made to: Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture.

Erratum: Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

RBP&#39;er er proteiner, der binder til det dobbelte eller enkeltstrenget RNA i celler og deltager i dannelse af RNA-protein-komplekser. RBP&#39;er kan binde en række RNA-arter, herunder mRNA og lange ikke-kodende RNA&#39;er (lncRNAs), og udøve deres indflydelse på post-transkriptionelle niveau. Både RBP&#39;er og lncRNAs dukker op som nye regulatorer i fedtvæv udvikling og funktion 1,2,3. For at forstå den mekanisme af RBP- og lncRNA-medieret regulering af cellulære veje, er det ofte nødvendigt at detektere interaktionen mellem en specifik RNA-molekyle og et eller flere RBP&#39;er, og undertiden at identificere det fulde spektrum af proteinpartnere af en RNA udskrift. Imidlertid kan eksperimentet være udfordrende på grund af det høje indhold af lipider i adipocytter. Den her beskrevne RNA pull-down protokol kan anvendes til at hente de proteinpartnere af en specifik RNA fra adipocyt lysater af primære kulturer 4,5. Baggrunden for denne protocol opsummeres som følger. Et RNA agn er in vitro transkriberet fra en DNA-skabelon, biotinmærket og konjugeret til streptavidin-belagte magnetiske perler 4. RNA agn inkuberes med cellelysater at tillade dannelsen af ​​RNA-protein-komplekser, der efterfølgende trukket ned på et magnetisk stativ. Mere specifikt RNA pull-down protokol beskrevet nedenfor anvender en RNA-fragment fra AR 3&#39;UTR som lokkemad for at hente Hur protein, et universelt udtrykte RBP bundet til 3&#39;UTR af mRNA&#39;er 6,7, fra theadipocytelysate af primær kultur. For at teste bindingsspecificiteten af ​​denne analyse, er en ikke-relevante RNAas samt tomme streptavidinkugler inkluderet som kontrol. Denne protokol er kompatibel med western blotting eller massespektrometri (MS) for at bekræfte erobringen af en bestemt RBP eller til at identificere den fulde repertoire af tilfangetagne RBP&#39;er henholdsvis 8. Flere teknikker er tilgængelige til at studere RNA-protein-interaktions. At afsløre RNA&#39;er bundet af en given RBP, RIP (RNA immunpræcipitation) og CLIP (UV tværbinding og immunpræcipitation) kan anvendes. I modsætning til at identificere protein partnere i en given RNA, RNA pull-down, kan anvendes Chirp (kromatin isolation af RNA oprensning), CHART (capture hybridisering analyse af RNA-mål) og RAP (RNA antisense rensning). I sammenligning med de senere dem, RNA pull-down teknik tager mindre indsats for at sætte op. Det kan anvendes til at indfange proteiner in vitro og in vivo. In vivo tilgang af RNA pull-down, som er teknisk mere udfordrende end dens in vitro modstykke, bevarer RNA-protein interaktioner ved tværbinding i celler, indfanger aptamer-mærkede RNA af interesse fra cellerne, og efterfølgende opdager bundne RBP&#39;er. RNA pull-down kan anvendes til at berige lave rigelige RBP&#39;er og at isolere og identificere de RNA-protein-komplekser, der har forskellige funktionelle roller i at kontrollere cellulær regulering 9,10 </sop&gt;.

<table border="0" cellpadding="0" cellspacing="0" width="857">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:499px;">Major materials used in this study.</td>
			<td style="width:177px;"></td>
			<td style="width:109px;"></td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MEGAscript kit</td>
			<td>&#160;Ambion</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biotin-14-CTP&#160;</td>
			<td>Invitrogen</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NucAway spin columns&#160;</td>
			<td>Ambion</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dynabeads M-280 Streptavidin&#160;</td>
			<td>Invitrogen</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HuR (3A2) mouse monoclonal antibody (sc-5261)&#160;</td>
			<td>Santa Cruz Biotechnology</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Goat anti-mouse IgG-HRP(sc-2005)&#160;</td>
			<td>Santa Cruz Biotechnology</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hypure Molecular Biology Grade Water (nuclease-free)&#160;</td>
			<td>HyClone</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate Buffer Saline (1&#215;PBS)</td>
			<td>&#160;HyClone</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RNase inhibitor&#160;</td>
			<td>Bioline</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Protease inhibitor&#160;</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pfu Turbo DNA polymerase&#160;</td>
			<td>Agilent Technologies</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TRIsure</td>
			<td>&#160;Bioline</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Name</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Major equipment used in this study.</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sorvall Legend Micro 21R centrifuge&#160;</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sorvall Legend X1R centrifuge&#160;</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Bio-Rad T100 thermal cycler&#160;</td>
			<td>Bio-Rad</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nanodrop 2000&#160;</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">BOECO rotator Multi Bio RS-24&#160;</td>
			<td>BOECO,Germany</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Protein gel electrophoresis and blotting apparatus&#160;</td>
			<td>Bio-Rad</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DynaMag-2 magnet&#160;</td>
			<td>Life Technologies</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

detection of rna-binding proteins by in vitro rna pull-down in adipocyte culture

RNA-bindende proteiner (RBP&#39;er) dukker op som en regulerende lag i udvikling og funktion af fedt. RBP&#39;er spiller en central rolle i genekspression regulering på posttranskriptionel niveauer ved at påvirke stabiliteten og translationel effektivitet af target mRNA. RNA pull-down teknik er blevet meget anvendt til at studere RNA-protein-interaktion, der er nødvendige for at belyse den mekanisme underliggende (lncRNAs) funktion RBP&#39;er &#39;samt lange ikke-kodende RNA&#39;er «. Men den høje lipid overflod i adipocytter udgør en teknisk udfordring i at gennemføre dette forsøg. Her en detaljeret RNA pull-down protokol er optimeret til primær adipocyt kultur. Et RNA-fragment fra androgen receptors (AR) 3 &#39;utranslaterede region (3&#39;UTR) indeholdende en adenylat-uridylate-rige elementwas der anvendes som et eksempel for at vise, hvordan man kan hente sin RBP partner, Hur protein fra adipocyt lystate. Den her beskrevne metode kan anvendes til at detektere interaktioner mellemRBP&#39;er og ikke-kodende RNA, samt mellem RBP&#39;er og kodning RNA.

I denne demo eksperiment blev en AR RNA-fragment anvendes som lokkemad for at fange dets bindende protein Hur. Både en FL RNA lokkemad, der er ikke-relevant for Hur protein og en alikvot af ukonjugerede datoer streptavidinkugler tjente som negative kontroller. RNA-protein-interaktioner kan forekomme enten i kernen eller cytoplasmaet, og dette pull-down protokol kan anvendes til enten total eller fraktioneret (nukleare eller cytoplasmatiske) cellelysater. Analyse af proteiner ved western...

Watch this Scientific Journal Video about Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture at JoVE.com

Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture

En RNA pull-down protokol er optimeret her til påvisning af interaktioner mellem RNA-bindende proteiner (RBP&#39;er) og ikke-kodende samt kodende RNA&#39;er. En RNA-fragment fra androgen receptor (AR) blev anvendt som et eksempel for at vise hvordan man kan hente sin RBP fra lystate af primære brune fedtceller.

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