Talk:Nucleofection

delete this article?

I think this article should specifically state how this method is different from electroporation, if it is, or it should be deleted and this should forward to electroporation. Are they just adding salt or something to the medium and pretending this is some revolutionary technology? When it is just electroporation tailored to a cell, that scientists have probably been doing for a long time? This isn't marketopoedia, and this article is all fluff and no substance as is.--Xris0 (talk) 04:53, 13 April 2010 (UTC)

but ih think there is some extra information, so it should be merged with electroporation and the name of nucleofection mentioned in that article, as it is a special form of electroporation for mammalian cells and primary cells, not bacterial transformation

Right. Please don't just delete. Redirect and merge with Electroporation.--Biologos (talk) 10:31, 13 April 2010 (UTC)

Is there anything to even merge? mammalian cells are transfected all the time without calling it nucleofection. I haven't electroporated mammalian cells but am guessing there are people out there who have and don't call it nucleofection, unless they're directly endorsed by this manufacturer. I request whoever wants to preserve anything here state what makes nucleofection unique/notable in clear terms.--Xris0 (talk) 18:12, 13 April 2010 (UTC)

Wait a minute. Are you really saying that you want to delete this article if you don't comprehend the difference between electroporation of bacteria and the delivery of DNA into the nucleus of a eukaryotic cell by electroporation by April 20? Because scientists have "probably" electroporated cells for a long time? They have not. The three referenced articles will give you some information. Another publicly available source I found in 1 minute is http://www.ncbi.nlm.nih.gov/bookshelf/br.fcgi?book=frsynapse&part=ch11#ch11.s11 . Believe me, I am as much against "marketopedia" as you are, but to delete information based on a hunch that it is redundant, and demanding clarification in 7 days, ist over the top, IMO. I have no time until April 20, but I promise that I will try to do the merge or to clarify the article in the next few weeks. Cheers, --Biologos (talk) 10:57, 16 April 2010 (UTC)

Well, I think this is Wikipedia and no sales promotion. The part 'Mechanism' contains no mechanism, and the whole article tells us nothing but that this method is wonderful and revolutionary and where a nucleofector can be buyed. There is absolutely no information in the article and all the links HOW nucleofection works, except that the result of this procedure is delivery of substances to the nucleus (not to cytoplasm as by standard electroporation procedures) by some electrochemical voodoo. Without this information (which I would expect from Wikipedia), this article is quite useless, except for AMAXA/LONZA's marketing. As I guess they will not talk about their patent-registered methods, nucleofection should be mentioned in the electroporation article at the best and the nucleofection article has to be deleted. --HStephan (talk) 09:23, 18 August 2010 (UTC)

Right, the method is patended. Therefore anyone can access the patents and extract information about the method to be used in this article. I have to admit that I did not keep my promise from April, though, and that I won't be able to do so in the next few weeks, either. If someone would take the information from this article (i.e. that there is an electroporation technique, called nucleofection, that can deliver nucleic acids into the nucleus of eukaryotic cells) and add it to Electroporation, the present article could really be changed into a Redirect. Alternatively, someone should add more information on the mechanism (for instance by looking at the patents) to this article.--Biologos (talk) 14:23, 18 August 2010 (UTC)

I think they they modified a few things in electroporation and got some extra efficiency,so they might have thought that it is best thing to get name and fame. —Preceding unsigned comment added by 220.225.126.138 (talk) 11:28, 6 December 2010 (UTC)

I think this should be merged with electroporation. As mentioned before, the mechanism is not explained at all and the whole article sounds like marketing. Some of the claims are questionable like all the statements about how only nucleofection efficiently delivers to the nucleus and so on. Microinjection for example also delivers to the nucleus and was invented over 100 years ago. Impalefection makes the same claim (more or less directly but at least with an explanation of the mechanism in the article).--Enblixt (talk) 13:26, 13 March 2012 (UTC)

My personal opinion: delete! It's nothing but a marketing trick by the company that sells "nucleofection". The company (Amaxa/Lonza) has never directly stated what is it their solution or their electroporator device that makes DNA magically go into nucleus. My educated guess however: 1) They sell 100% standard electroporator not much different from any of their competitors, 2) the "nucleofection" part is achieved by adding a DNA-binding nuclear localization signal peptide to the electroporation solution (and that is why they sell the solution without ever disclosing what it contains). And if I had to bet, it's a peptide from the SV40 virus Large T Antigen as described in this paper long before "nucleofection": http://www.ncbi.nlm.nih.gov/pubmed/3015419 Merging it with electroporation would be wrong because it is NOT a special form of electroporation! — Preceding unsigned comment added by 128.104.117.239 (talk) 19:54, 7 May 2012 (UTC)

I finally found the patent, with patent numbers including US 2004/0137622 A1, US 7320859 B2 and EP 1427837 B1 (see for instance ip.com/patapp/US20040137622 ). The abstract says:

The present invention relates to a method for transfection of cells using at least one protein capable of forming nucleoprotein filaments, wherein the protein is initially modified with at least one functional component which influences one or more steps of the transfection, the nucleic acid to be transfected is then loaded with the modified protein, whereby the nucleic acid and the protein form a filament-like complex, and this complex is finally added to the cells to be transfected.

I haven't had time to thoroughly read it, but the trick seems to be the combination of a DNA-binding filamentous protein like RecA with a nuclear localization sequence, and maybe another protein sequence to aid in binding to the cell surface before electroporation.

I also found a patent for their electroporator, but I don't understand enough of it to say whether it is just a standard electroporator: US 2006/0094095 A1, see for instance www.google.com/patents/US20060094095.pdf --Biologos (talk) 10:44, 8 May 2012 (UTC)

Re: "the trick seems to be the combination of a DNA-binding filamentous protein like RecA".

Don't think so. More likely, it just means that they decided to try something else and patented it 'just in case'. Why not spike DNA with a tiny bit histones instead? (It works, I know, I tested but never bothered to publish). That same patent cites Amaxa's patent on NLS peptides - and peptides seem to work just as well as proteins (be it RecA-like things or just histones). Peptides are cheaper and much more stable. Besides - and this is critical argument - they now sell "nucleofection PLUS supplements" which you can add to cells for freezing and storage before electroporation. These supplements are to be used in place of the regular supplements. Which means 1) they contain high DMSO concentration, 2) they contain Amaxa's magic nucleofection additives. Ergo, the additives are super likely to be peptide(s) rather than proteins because the latter would not be stable in high DMSO concentration. As for electroporator, the one that they sell as absolutely standard one, only distinguished by pre-programed methods loaded into memory and the one described in the patent is also very standard one but containing pretty elaborate and, frankly, ridiculously unnecessary safety and arcing prevention features. — Preceding unsigned comment added by 68.117.135.21 (talk) 03:11, 13 May 2012 (UTC)

removing POV tag with no active discussion per Template:POV

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Since there's no evidence of ongoing discussion, I'm removing the tag for now. If discussion is continuing and I've failed to see it, however, please feel free to restore the template and continue to address the issues. Thanks to everybody working on this one! -- Khazar2 (talk) 01:14, 30 June 2013 (UTC)