Transcription and Translation

RNA + Body Fluids (the results are in)

OK I finally did that experiment that people asked for … and more.

If you want to know why I performed this experiment, read this post on RNA treatments for autism. Here I present to you evidence that your body is secreting enzymes, called RNAses, that will chew up your RNA in minutes.

So the experiment:

500ng of RNA was mixed with either
– control buffer (100mM KCl, 10mM Tris buffer pH7.4)
– 1ul of DNAse (1mg/ml), this is what you would call a negative control – no degradation should be seen here
– 1ul of water applied to sweaty skin
(After working a bit with my latex gloves, I puled off my gloves and applied 20ul of buffer for 5 secs and then collected the drop of sweat+buffer sample)
– 1ul of saliva (collected from a baymate)
– a single strand of hair (again collected from my trusty baymate)
– a 5mm long toe nail shaving (again from my baymate)
– 1ul of RNAse (1mg/ml), this is a positive control – we should see plenty of RNA degradation here

(note ng = nanogram; ul = microliter = 1/1000 of a milliliter)

All samples were brought up to 5ul total volume using the control buffer and incubated at 37 degrees Celsius for 5min. Note that the RNA is present at 0.1mg/ml, a decent concentration.

The reaction was terminated by adding 5ul of 50% formamide loading buffer and then incubating the samples at 90 degrees Celsius for 10min.

Samples were loaded on a prestained non-denaturing agarose gel (I didn’t have time to make radioactive RNA and pour a denaturing gel) and a 90mA current was applied to the sample for 30min.

And the result:

i-f85f2d5a0e627543d4a954b45049de7e-bodyRNAse.jpg

The RNA migrated from top to bottom. The longest RNA strands migrated the slowest and are thus on the top of the gel. Smaller RNA strands migrated faster are thus on the bottom. To show you how fast the bands migrated, I loaded molecular weight standards in the first lane. 2kb = 2 kilobases = an RNA strand that is 2000 bases long. The substrate for this reaction was an RNA species that I synthesized in the lab. This transcript is 500 base pairs long. It was capped at the 5′ end and polyadenylated (i.e. a poly-A tail was added) to the 3′ end. There is about 200-300 adenines in the poly-A tail and so the whole RNA is about 700 to 800 bases long. This is a good mimic for the majority of the mRNA inside a cell (actually this particular transcript is fairly stable compared to the typical mRNA transcript). Although the samples were loaded on a non-denaturing gel, this RNA tends to run at the proper molecular weight as it does not form much secondary structure.

As you can see,
DNAse had little effect, the RNA is still there and migrates at the right speed.
Sweat can degrade RNA. So despite the fact that this reaction was carried out with water that came into contact to a sweaty hand, the end result was that the RNA was effectively chopped into bits after a simple 5 min incubation. The size of the bits are about 10-100 bases long.
Saliva can degrade RNA very effectively. Don’t forget that this sample’s saliva is 5 times less concentrated than what is in your mouth. Some small RNA bits survived the treatment (maybe 10-50 bases long) but the majority of the RNA has been decomposed to smaller stuff (single nucleotides) that can’t be stained.
The single strand of hair had minor effects. Actually I quantitated the intensity of the band and it’s only 80% of the control – so as much as 20% of the RNA was affected, but the majority of the RNA survived.
Toe nail clippings were deadly to the RNA. Lesson: don’t try to salvage RNA that landed on your toe nail!
– pure RNAse was … well lets say that I added the equivalent of a nuclear bomb to the poor RNA. Not even a trace survived.

So there you have it. We are all oozing RNAse, the enzyme that degrades RNA. Stick RNA in your kid’s mouth and it’ll be gone – digested by the RNAse found in human saliva. And there’s a whole lot more RNAse in your stomach – I promise you that no RNA will ever reach your intestines (where all the nutrient absorption takes place). If you read my previous post on RNA & autism, you’ll find out that “therapeutic” RNA is sold in lan aqueous solution with a dripper. Even touching the dripper your bare hands is enough to degrade all the RNA inside the vial in minutes. The fact that your body is full of RNAse is not surprising as our bodily secretions help protect us from many viruses that have RNA genomes.

Conclusion:
Direct RNA ingestion will never work.

Acknowledgements:
I would like to thank baymate for suplying bodily fluids and parts.

Comments

  1. #1 lazybratsche
    November 29, 2006

    Don’t you wish that every experiment was this easy, conclusive, and entertaining?

    Great work btw. I spent one summer in an RNA lab, so I can understand the absolute shock and horror conjured by the idea of putting RNA in someone’s mouth.

  2. #2 Kathleen Seidel
    November 29, 2006

    You are a man of your word! Thanks so much for doing this, and thanks to baymate, too.

  3. #3 apalazzo
    November 29, 2006

    I reread this and it’s full of typos. Sorry about that, I’ll clean it up.

    PS I’ll pass on the good words to the baymate.

  4. #4 Terry
    November 29, 2006

    Mind if I use that in one of my lab class, with a link to this post?

  5. #5 SMC
    November 29, 2006

    How long before the phrase “RNA Treatment” gets replaced by “Nucleotide Supplementation” in the crackpot literature for exactly the same magic anti-autism pills?…

  6. #6 Orac
    November 29, 2006

    Actually, I’m surprised the RNA didn’t degrade even more.

    One unexpected result: I would never have expected that a toe nail shaving would be more potent at degrading RNA than saliva. That’s some nasty shit under those toes.

  7. #7 Theodore Price
    November 29, 2006

    Hey, lets be nice to his baymate :-). One of my toenail clippings would have given RNase nuclear bomb results.

    So any plans to post this in the forum on the autism answers site? That would be a riot!

  8. #8 Bartholomew Cubbins
    November 29, 2006

    Given that your lab is funded, in part, by the NIH, it is obvious that you have an agenda to shackle the righteous and true supplements and alternative therapy industry. So sayeth Orrin Hatch. Is a sarcasm tag even necessary?

    Unless I missed it, you might want to make sure that people know where you’re getting the RNA from – total RNA extract from yeast? That’s what they’re forcing down their kids’ gullets.

    Theodore – they already know, that’s the sad part.

    Alex, I saw some funny comments in a different blog remarking that the alties have a real penchant for anal suppositories. So I have to ask, how far will your benchmate go for the sake of science? I’m thinking that this will work: an EtOH ppt of a total RNA extract followed by drying the pellet and putting the powder into an empty tylenol capsule shell. Add one part UFIA and presto! I’ll leave out the gory details, but your benchmate could even do a timecourse and then post that gel to Amy’s fanfic site.

  9. #9 Dixon Ng
    November 29, 2006

    Great experiment Alex! The simplicity of it illustrates the elegance of science :)

  10. #10 Dixon Ng
    November 30, 2006

    PS – If you ever swing by Vancouver BC, let me buy you a drink.

  11. #11 apalazzo
    November 30, 2006

    Mind if I use that in one of my lab class, with a link to this post?

    No problem.

    One unexpected result: I would never have expected that a toe nail shaving would be more potent at degrading RNA than saliva. That’s some nasty shit under those toes.

    RNAse is pumped into your saliva, in contrast toe nails just picks up stuff from the environment.

    TP – I’ll send this to the next Skeptics’ Circle which is being hosted by an Autism guy.
    BC – I’ll bring it up with my baymate …
    NG – I may take you up on that. My inlaws are in Seattle so we tend to travel to that side of the continent once a year. + I really like Vancouver.

  12. #12 jim
    December 22, 2006

    Hi Dr. Amy,
    I guess it has become known that I am following your protocol and, as a physician and longtime member of some of the talk groups, several parents have asked me how the RNA works. They seem to be insisting that after investigating it, that it is a homeopathic formula with RNA imprints that vibrate with the frequency of the RNA’s. I have told them that this is not at all my understanding of what is in the RNA bottles. Can you put this idea to rest and answer those who say that the RNA is just homeopathy? I think that is important because there is a lot of misunderstanding and confusion about this.
    Thanks.
    Ken

    >> Dear Ken,

    The long answer is that they should read the RNA book and listen to the
    tapes. It took me 200 pages to explain how RNA works it would be
    impossible to do it in a short email.

    However, I do know what you are asking…RNA has been used as a
    nutritional supplement, a homeopathic drug and a medical food. In all of
    these cases you are using a total RNA extract. This will in fact provide
    some benefit and there is a long list of literature that supports it’s use
    for many years. Taking advantage of my background in biotechnology I am
    able to bring a fresh approach to this field. What I am doing is a blend
    of the best of the natural approach combined with the specificity of the
    biotechnology approach. I am able to do this aft er hav ing spent 20years in
    biotechnology as well as being trained in the use of natural supplements.
    The RNAs that I use in my practice are from yeast RNA (contain NO actual
    yeast). They are specific combination of RNAs that represent a distinct
    advantage over total RNA mixtures. As discussed in Arizona, it
    is expensive to isolate a sinlge RNA or even 10 to 25 RNAs from a total
    mixture of 1 million RNAs. This is very different from using a total
    homeopathic RNA, and again as discussed a total RNA mix will not be nearly
    as effective. If you refer the questioner to the powerpoints that are on
    this site from the RNAs talks, at the end of the Understanding RNA talk I
    went through the math. This is the slide that looks at the total number o f
    m illigrams in a total yeast RNA capsule, divides by the number of
    molecules to show that if you are using 10 to 25 of the yeast RNAs of
    choice that it will take the number of micrograms that are in each bottle
    to do this. This is not just vibrational frequency.

    HEEL is one of the more respected Homeopathy companies. The medical
    director of HEEL was on TV last week talking about using the RNAs that I
    have developed and how they have changed his life. I think this speaks to
    the enhanced value of these RNAs.

    Again Ken, there are a lot of people who will continue to insist that the
    world is flat until there can see it from another angle. I expect that the
    ones asking the questions have not read the RNA book, have not watched the
    videos, and have not tried these RNAs. I am not sure that it matters what
    you say. They are threatened by something new that they cannot u nderst an d
    and may not be willing to try to even understand it.

    If I wear a red and green suit the next time that I give a talk, you will
    be able to sit in the audience and see my red and green suit. On the other
    hand if one of the individuals asking you the RNA questions is in the
    audience and happens to be color blind, he or she will look at me and see
    a brown suit. No matter how hard you try to explain that the suit is red
    and green and that you can clearly see it, they will still see brown.
    Until they are willing to acknowledge that even if they can’t see it, it
    may be real, you will be frustrated with them.

    My approach is to understand and accept them, and to appreciate that they
    are not yet ready to embrace something new and exciting that can make a
    profound diffence in healthcare.

    Finally, many of those casting stones also suggest that the RNAs are
    expensive just as a way to tak e adva nta ge of autistic parents. As I have
    already explained it is very expensive to isolate the “needles out of a
    haystack” in terms of using only those yeast RNAs of choice. I also
    seriously doubt that there is another doctor out there who donates
    almost 30 hours a week for free, working 7 days per week to answer parent
    questions out of their personal time.

    I thank you and send you my love for trying,
    Amy

  13. #13 Bartholomew Cubbins
    December 27, 2006

    Hey Alex – this is hilarious. Assuming this is real, kudos to whoever posed this question to her and got her to actually write this.

  14. #14 Dixon Ng
    December 29, 2006

    That’s a good post jim. No wonder we couldn’t figure it, its homeopathy! *As an aside, anyone interested in the ‘principles’ of homeopathy should really watch James Randi’s Princeton lecture (just google it).

  15. #15 Alex
    May 5, 2007

    Hi Alex,

    I found out about your blog as a result of your article on RNA treatment of autism. I thought it was very educational and interesting. I know some who are using the RNAs to give to their autistic children. I will have to tell them about your article and experiment and see their response.

    I know this is not your specialized field, but I wanted to know what you think of the Titers test. My older son is on the autistic spectrum and our allergy/immunology doctor recommended that we take this test. The results showed an off-the-chart score relating to Rubella and a higher than normal score relating to Mumps and Measles.

    Do you think this means anything? Is it a legitimate test and if so, what do his high scores indicate, especially in relation to Rubella, where it was literally off the chart?

    Also, what do you know about QEEGs (a.k.a brain maps)?

    Please let me know.

    Thanks,
    Alex

  16. #16 skeptic
    June 20, 2007

    I enjoyed your experiments with RNA. I have been spying on Yasko’s board and am appalled by the pseudo scientific mumbo jumbo she spouts about her supplements (you just need an email address to join: http://www.ch3nutrigenomics.com/phpBB2/index.php). Of course she always has four possible responses to whatever people post: 1) increase your dosage, 2) add more supplements, 3) give it more time, or 4) all of the above.

    You may have seen the website or references to those who scam the Nigerian email scammers (you know, the email that asks for a bank account # so you can share millions with them). It is quite funny to see the scammers get suckered. Has anyone with the right bio-chem credentials thought of doing the same with Yasko? Inventing ailments and asking for advice in a way that would expose the lack of science and contradictions in her program? I would be happy to help!

  17. #17 Ann
    August 24, 2007

    Thanks for doing this! As the mother of a child with autism, I’m investigating different theories and interventions. Another parent recommended Dr. Yasko, and my search led me here. I wanted to ask if you tested the RNA supplement brand that Yasko sells or promotes or if you tested RNA from your own lab. I imagine Dr. Yasko would say that her endorsed supplement is stabilized somehow and remains effective, unlike “common” RNA. :) Thanks, again.

  18. #18 apalazzo
    August 24, 2007

    Dr Yasko claims that her RNA is extracted from yeast. In addition Dr Yasko sells her product as a supplement and not as an FDA approved drug and thus by law she can’t chemically modify it once it is extracted. If you read her literature carefully she never claims that her RNA is modified, only that RNA can be modified to make it more stable. In other words, this product only contains unmodified RNA, and thus will behave like the RNA used in the above experiment.

    Even if the RNA did survive all the enzymes in your child’s saliva, her claims that RNA can act as a chelator are totally unfounded. This product is essentially a placebo.

  19. #19 Jes
    April 22, 2010

    Simple yet proves the point experiment. Good job. I hope you get a tenure.

  20. #20 Uber Chuchem
    April 27, 2010

    RNA Therapeutics Workshop brings together medical community in Lisboa:

    http://www.hmsportugal.org/NewsDetail.aspx?contentId=160
    ” In the academic field Michael Czech from the University of Massachusetts discussed some of the most promising systems for oral RNA delivery based on the use of baker’s yeast cells.”

  21. #21 alena
    March 21, 2012

    As a biochemist (PhD) and a single mom of 2 kids with autism. I find Dr. Yasko and most of the biomedical community appalling. I had a mom tell me “hate the autism not the child” sigh….but i Love ALL of my children..even the autistic part..i stopped thinking of them as autistic a long time ago and started thinking of them as who they are..2 kids….

    I have a problem with so many of these biomedical treatments..do you know they give CHARCOAL supplements..to “get out the bad yeast” the charcoal binds to the bad yeast toxins so you’re not so yeasty..

    a friend of mine (her kids have autism as well) always joke about the fact that we are yeasty…it’s nice to see real researchers look into this! :)

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