tag:blogger.com,1999:blog-45514594291597358642024-02-20T00:20:32.029-08:00Claudiu D. Ursachi blogWelcome to my blog about induced pluripotent stem cells (iPSCs)Claudiu Daniel Ursachihttp://www.blogger.com/profile/01154854650933573325noreply@blogger.comBlogger2125tag:blogger.com,1999:blog-4551459429159735864.post-20392629054248663092010-09-16T13:07:00.000-07:002017-11-29T13:26:44.909-08:00Last updated: September 6, 2017.<br />
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<b><span style="color: #000099;">iPSCs blog</span></b><br />
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<b><span style="color: #000099;">Contents</span></b><br />
<span style="color: #000099;">(Select an underlined item)</span><br />
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<span style="color: #000099;">Unit One:</span> <span style="color: #3333ff;"><b>News induced pluripotent stem cells (iPSCs)</b></span><br />
<span style="color: blue;">- </span><a href="http://claudiuursachi.blogspot.ca/p/news-induced-pluripotent-stem-cells.html" target="_blank"><span style="color: blue;">2011</span></a><br />
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<span style="color: #3333ff;"><b></b></span><span style="color: #3333ff;"><span style="color: #000099;">Unit Two:</span> <b>My articles about induced pluripotent stem (iPS) cells</b></span><br />
<span style="color: #3333ff;"><b>-</b> </span><a href="http://claudiuursachi.blogspot.com/2010/07/episomal-expresion-of-lin28-proteins-in.html"><span style="color: #3333ff;">The Episomal Expression of the Protein Lin28 in Mammalian HEK 293 6E cells</span></a><br />
<span style="color: #3333ff;">- </span><a href="http://claudiuursachi.blogspot.com/p/structures-and-some-interactions-of.html"><span style="color: #3333ff;">The Structures and Some Interactions of the Proteins Oct-4, Sox2, NANOG and Lin28 That Are Used to Induce the Pluripotent Stem (iPS) Cells</span></a><span style="color: #3333ff;"><br /></span><br />
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<span style="color: #993399;">Unit Three: <b>Characteristics of induced pluripotent stem (iPS) cells</b>- </span><a href="http://claudiuursachi.blogspot.com/p/different-cells-used-for-reprogramming.html"><span style="color: #993399;">Different cells used for reprogramming </span></a><br />
<span style="color: #993399;">- </span><a href="http://claudiuursachi.blogspot.com/p/characteristics-of-induced-pluripotent.html"><span style="color: #993399;">iPSCs derivation methods</span></a><span style="color: #993399;"><br />-</span><a href="http://claudiuursachi.blogspot.com/p/molecules-controlling-pluripotency-in.html"><span style="color: #993399;">Molecules</span><span style="color: #993399;"> that control the pluripotency in embryonic stem (ES) cells and induced pluripotent stem cells (iPSCs)</span></a><span style="color: #993399;"><br /></span><span style="color: #993399;"><br /></span><br />
<span style="color: #993399;">Unit Four: <a href="http://claudiuursachi.blogspot.com/p/bioinformatic-analysis-es-and-ipscs.html"><b><span style="color: #993399;">Bioinformatic analysis ES and iPSCs</span></b></a></span><span style="color: #993399;"><b><br /></b></span><br />
<span style="color: #993399;"><b><a href="http://claudiuursachi.blogspot.ca/p/my-resume.html" target="_blank">My Résumé</a></b></span><br />
<span style="color: #993399;"></span><br />Claudiu Daniel Ursachihttp://www.blogger.com/profile/01154854650933573325noreply@blogger.com0tag:blogger.com,1999:blog-4551459429159735864.post-34686546767574816402010-07-10T14:44:00.000-07:002017-11-29T07:15:11.559-08:00The Episomal Expression of the Protein Lin28 in Mammalian Cells HEK 293 6E<div align="justify">
C.D. URSACHI<br />
Dec. 2009<br />
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<span style="color: #3333ff;"><b><b>Abstrac</b>t</b> </span><br />
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The protein <a href="http://www.pdb.org/pdb/explore/explore.do?structureId=2CQF">Lin28</a> and <a href="http://www.nature.com/ncb/journal/v11/n3/abs/ncb0309-228.html">miRNA</a> <a href="http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6TCX-4TCCHT0-1&_user=10&_coverDate=10%2F31%2F2008&_rdoc=1&_fmt=high&_orig=browse&_origin=browse&_sort=d&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=323f053fe8f5e50513d43cb91607deef">Let-7</a> form a cellular control <a href="http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6W7J-4T4108X-1&_user=10&_coverDate=09%2F30%2F2008&_rdoc=1&_fmt=high&_orig=browse&_origin=browse&_sort=d&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=d2efeaf4ffe9f8084ef381a0a37b88a8">system</a> that is an important regulator of gene expression that controls both physiological and pathological processes such as development and <a href="http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T23-4N14677-1&_user=10&_coverDate=06%2F30%2F2007&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_searchStrId=1446812955&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=b4941861db31cb2646e138c94c572171&searchtype=a">cancer</a>. Firstly, the miRNA Let-7 has been reported to play a <a href="http://www.jbc.org/content/283/31/21310.long">tumor suppressor role </a>by repression of some oncogenes (<a href="http://www.ncbi.nlm.nih.gov/gene/8091">Hmga2</a>, Ras, and <a href="http://www.ncbi.nlm.nih.gov/gene/4609">Myc</a>). Secondly, Lin28 is a protein that <a href="http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2758923/pdf/nihms-140663.pdf">blocks maturation of Let-7</a> and it is reported among the factors that reprogram human somatic cells to pluripotent stem cells (<a href="http://www.gs.washington.edu/news/Yu.pdf">iPS</a>). By adding, that <a href="http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6WSR-4TR9CHY-H-2&_cdi=7053&_user=10&_pii=S1097276508006606&_coverDate=10%2F24%2F2008&_sk=%23TOC%237053%232008%23999679997%23700024%23FLA%23display%23Volume_32,_Issue_2,_Pages_159-300_(24_October_2008)%23tagged%23Volume%23first%3D32%23Issue%23first%3D2%23date%23(24_October_2008)%23&view=c&_gw=y&wchp=dGLzVzz-zSkWb&md5=41d661c786b57ecc681f2eca30fafaab&ie=/sdarticle.pdf">Lin28 is highly expressed in undifferentiated cells</a> and certain cancers, it could be an interesting protein for the <a href="http://www.ingentaconnect.com/content/mksg/pto/2005/00000096/00000003/art00002">development of the drugs</a>. In this context, the purpose was to verify the episomal expression of the protein Lin28 in human embryonic kidney (HEK293-6E) cells despite the interactions between Let-7 and Lin28 that determine several embryonic cell transformations. To verify the production of recombinant protein (r-protein) Lin28, we used a system of r-protein production form by HEK293-6E cells (suspension culture), recombinant vector (pTT5SH8Q1-Lin28) with cytomegalovirus (CMV) promoter, linear polyethyleneimine (PEI) as transfection agent and the culture medium serum-free (<a href="http://cshprotocols.cshlp.org/cgi/content/full/2008/3/pdb.prot4977?maxtoshow=&hits=10&RESULTFORMAT=&fulltext=Transfection+of+HEK293-EBNA1+Cells+in+Suspension+with+Linear+PEI+for+Production+of+Recombinant+Proteins&searchid=1&FIRSTINDEX=0&resourcetype=HWCIT">P</a>). After the construction of recombinant vectors (pTT5SH8Q1-Lin28), we verified the production of Lin28 using three ratios DNA: PEI and two harvests of HEK293-6E cells (24h and 48h). The verification of the expression of Lin28 has been carried out by SDS-PAGE electrophoresis. The protein Lin28 was synthesized in all experimental conditions, but production was greater after 48h after transfection and a ratio DNA: PEI of 1:2. In conclusion, the protein Lin28 has been produced by episomal expression in HEK293-6E cells (suspension culture) using a recombinant vector (pTT5SH8Q1-Lin28).<br />
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<span style="color: #3333ff;">Correspondence</span>: claudiu.ursachi@videotron.ca</div>
Claudiu Daniel Ursachihttp://www.blogger.com/profile/01154854650933573325noreply@blogger.com0