This is worrying if true....

philbar72

Gene doping, naturally produce (sic) more red blood cells. Never seen this before, and don't know if it defeats the biological passport tests!

http://www.cyclingtips.com.au/2013/02/t ... ne-doping/

ddraver

We need a biochemist for this, but I don't understand why you can't test that cells don't have 2 genes for EPO....

fastandfurry

ddraver wrote:

We need a biochemist for this, but I don't understand why you can't test that cells don't have 2 genes for EPO....

You could either have got the right combination of genes (it's a lot more complicated than one gene for most things, imagine this is the same..) for producing more EPO through a fortunate combination of parents and luck (that you got the right particular set of genes from them), or from using gene therapy - how're you going to tell the difference just by looking at the end result/testing of DNA?

ddraver

that's why we need a biochemist - I thought that was pretty clear really by my use of the words "We need a biochemist for this"

inkyfingers

ddraver wrote:

We need a biochemist for this

I Told you you that geology degree was the wrong decision.

Richmond Racer

ddraver wrote:

that's why we need a biochemist - I thought that was pretty clear really by my use of the words "We need a biochemist for this"

Dont worry, there'll be one along any minute. In fact, on this forum, it'll be like buses...

skylla

The article on cyclingtips only briefly deals with genetherapy, but mainly focusses on HIF -hypoxia inducing factor - which is/are a set of protein factors that stimulate the cellular and physiological response to hypoxic conditions i.e. low concentrations of oxygen at for instance altitude. They are saying that the next PED is more likely to be a HIF than gene therapy as there are many as yet unresolved issues with the latter.

HIF can be detected in similar ways as EPO, they are both proteins, however because HIF needs to be in the cell for it to act (unlike EPO) it requires a 'vector' which, in principle, might make it even easier to detect. I also dare say that the glow time might be longer. In the end, as HIF will have a similar effect on reticulocytes/erythrocytes ratios as EPO, the biological passport will pick it up too.

RichN95.

I've read that this stuff is really easy to test for.

esafosfina1

Gene doping has been mooted for a few years now... as per usual there's rumours out there...

ddraver

inkyfingers wrote:

ddraver wrote:

We need a biochemist for this

I Told you you that geology degree was the wrong decision.

Yeah, well when we do the why are the roads in the Pyrenees steeper than the Alps question in July, I ll be there with bells on!

skylla

fidbod

ddraver wrote:

We need a biochemist for this, but I don't understand why you can't test that cells don't have 2 genes for EPO....

Now - bear in mind that I am relying on my ten year old Biology degree here... My initial thought is that testing for a second EPO gene is possible but impractical.

From the human genome project we know where the naturally occurring EPO gene is located i.e. its rough location on one of the 23 pairs of human chromosomes.

A second - artifically inserted - EPO gene could in theory be inserted onto any of 22 pairs of chromosomes (excluding the X/Y pair). So to tell if an athlete has two EPO genes would require sequencing the athletes entire genome and looking for multiple instances of the gene.

Not impossible but time consuming - you might be able to do it in a couple of days currently but whether that would generate data substantive enough to pass the evidence tests required for doping control is another matter....

a completely separate thought - proving that the second EPO gene is artificial rather than a product of natural human mutation would be tricksy as well.

skylla

fidbod wrote:

ddraver wrote:

We need a biochemist for this, but I don't understand why you can't test that cells don't have 2 genes for EPO....

Now - bear in mind that I am relying on my ten year old Biology degree here... My initial thought is that testing for a second EPO gene is possible but impractical.

From the human genome project we know where the naturally occurring EPO gene is located i.e. its rough location on one of the 23 pairs of human chromosomes.

A second - artifically inserted - EPO gene could in theory be inserted onto any of 22 pairs of chromosomes (excluding the X/Y pair). So to tell if an athlete has two EPO genes would require sequencing the athletes entire genome and looking for multiple instances of the gene.

Not impossible but time consuming - you might be able to do it in a couple of days currently but whether that would generate data substantive enough to pass the evidence tests required for doping control is another matter....

A few points:

** DNA sequencing is not impractical at all. However, testing for "gene therapy" will more likely involve a technique called southern and/or northern blotting, which is the use of a DNA probe to anneal to RNA/DNA sequences of your liking.

** An entire human genome did indeed take weeks, however it can now be sequenced in 30 minutes if so desired.

** Gene therapy only targets a subset of cells, i.e. it is tissue specific thus all one have to do is sequence comparison. There is no way gene therapy would affect all cells of a human body (unless we are talking designer babies, but that's a different discussion!)

** All mammals including humans have diploid cells, i.e. there's a gene from your mum and one from your daddy. The recombinant/exogenous EPO or HIF gene is thus the third and very likely to have exogenous flanking sequences that are non-human and can be easily, very easily picked up.

fidbod wrote:

a completely separate thought - proving that the second EPO gene is artificial rather than a product of natural human mutation would be tricksy as well.

*** "Artificial genes" are even easier to detect!

Garry H

inkyfingers wrote:

ddraver wrote:

We need a biochemist for this

I Told you you that geology degree was the wrong decision.

I have an Economics degree. Is this method financially viable?

dortmunder

No convinced this would work. Gene's are only switched on in certain cells and organs so to take over a group of cells or organs to produce more EPO would be practically impossible. I guess one way would be to use stem cells to produce cells capable of more EPO production but again the cost and lack of conclusive proof about stem cells would make it prohibitively expensive

skylla

dortmunder wrote:

No convinced this would work. Gene's are only switched on in certain cells and organs so to take over a group of cells or organs to produce more EPO would be practically impossible. I guess one way would be to use stem cells to produce cells capable of more EPO production but again the cost and lack of conclusive proof about stem cells would make it prohibitively expensive

It does work and is not impossible - it is routine work every single day for many a molecular biologist in experimental settings, from organisms as small as bacteria to as big as mice, sheep, cows, etc. Never mind the millions of human cell lines that are cultured in controlled conditions and used in experiments every single day. For us human beings, gene therapy is in advanced clinical trials. Also, you do not need stem cells - genes can be switched on by for instance genetic engineering, small molecular compounds or mimicking environmental conditions (HIF example).

PS: every living thing (bacteria, yeast, hamster cells) can produce EPO through a bit of engineering: it's a gene product!

frenchfighter

http://www.dailytech.com/MuscleGrowth+G ... e17082.htm

rob churchill

Mad_Malx posted a link to a technical article on dope testing about ten days ago in the "Frank Schleck tests positive for Diuretic" thread. It was mostly about conventional blood doping, but did include this:

In addition, in vivo Epo gene transfer could probably be detected if applied by athletes, as an IEF study revealed unusual Epo glycosylation forms on allogeneic Epo transfer into skeletal muscle of cynomolgus macaques via adeno-associated virus.72 In the initial studies of adeno-associated virus-mediated allogeneic Epo cDNA transfer to macaques, severe anemia developed in many animals after a few months, which was probably the result of an immune reaction.73,74 However, in using a rapamycin dimerizer-regulated gene expression system, Rivera et al75 achieved controlled, long-term production (up to 6 years) of Epo in rhesus monkeys, with no apparent immune response. Regarding the possibility of Epo gene doping in humans, strategies are under development to specifically amplify intron-less DNA sequences and PCR protocols allowing the detection of small amounts of transgenic DNA in blood.76–78 The tests take into consideration that transgenes are usually derived from the cDNA for the gene to be transferred and cDNA does not contain introns [...] In conclusion, Epo gene transfer is possible but medically little explored with respect to efficacy, safety, and immunogenicity. It seems less likely that any of the techniques has entered the sports scene.

They don't think medical gene therapy is yet at the point where athletes might abuse it, but they think it should be detectable if they ever do (and I would imagine, detectable for the rest of the athlete's life, which has got to be a disadvantage.)

skylla

1) Nice, but engineering strategies could use genomic DNA (with introns) with the same ease as cDNA (without introns), thereby bypassing detection. Either way, it would be so much easier to detect recombinant adeno-vector sequences!

2) The rapamycin experiment requires a previous round of genetic engineering in which a heterologous gene expression cassette is incorporated into the DNA of a cell. This is incredibly easy to detect, with a wealth of technologies developed in the sixties and seventies.

KieranHardman

I wouldnt worry about gene doping. If you could artificially insert a working series of genes into a mammal and make it work efficiently and routinely then you could cure so many fatal illnesses. That is far more financially beneficial than doping some athlete. Probably easier to get funding for as well and scientists bloody love funding. Nobel Prize for Medicine is far more desireable than having a customer win the Tour.

There was a paper I remember reading for one of my finals exams where an artificial gene was vectored into a number of human test subjects to counteract a potentially fatal disease. It worked in only a small number of cases. It is notoriously difficult to get the required section of DNA inserted not only in the right place, but in the correct reading frame and even in the right direction. This was in 2006 though so things may have moved on since then.

As for people doing it routinely now in animals. It happens but you are doing it at embryonic level when you can pick and choose which cells it works in. Cant do that with humans.

Source: Genetics degree from a while ago.

skylla

KieranHardman wrote:

I wouldnt worry about gene doping. If you could artificially insert a working series of genes into a mammal and make it work efficiently and routinely then you could cure so many fatal illnesses. That is far more financially beneficial than doping some athlete. Probably easier to get funding for as well and scientists bloody love funding. Nobel Prize for Medicine is far more desireable than having a customer win the Tour.

There was a paper I remember reading for one of my finals exams where an artificial gene was vectored into a number of human test subjects to counteract a potentially fatal disease. It worked in only a small number of cases. It is notoriously difficult to get the required section of DNA inserted not only in the right place, but in the correct reading frame and even in the right direction. This was in 2006 though so things may have moved on since then.

As for people doing it routinely now in animals. It happens but you are doing it at embryonic level when you can pick and choose which cells it works in. Cant do that with humans.

Source: Genetics degree from a while ago.

1) Inserting genes is relatively straight forward, even in 2006. The 'reading frame' and 'direction' is of no importance at all as you're not targeting a gene sequence, but an 'open' locus to drop your gene of interest. However, getting the damn things expressed is incredibly difficult. Mammalian cells are very efficient in 'silencing' foreign and ectopic DNA. It requires quite a bit of engineering to get gene expression levels right even with technologies available today in 2013.

2) There's a lot of money in sport and a lot of charlatan scientists. I don't know how well you're following medical advances in gene and stem cell therapies, but they are on offer in certain ex-soviet republics and of course china.

The financial rewards in running a gene doping clinic are of course much bigger than doing science! Whatever Fuentes was charging per blood bag is what a scientist can only dream of as a 'reward' for a month's work. There's a lot of sportsmen who are willing to spend hundreds of thousands on unproven gene and stem cell therapies and this is already happening.

The Nobel v. TdF argument really doesn't hold any water as there is only 1 prize in medicine a year and the one for discovery of pluripotent cells (i.e. stem cells) has already been given away in 2012. The prize for recombinant DNA was given away in 1980! There's a lot more prizes a year in professional sports!

EPO Delivery Man

Sounds like b0llocks to me and even if it was true, its only a matter of time before the Taiwanese get their hands on it and we have a lot of 'fake genes' knocking about...


come on..that wasn't bad eh..and im in work..

Macaloon

Bonus Kimmage-Cam footage from the Tyrell Corporation inquiry.
Jens meets his maker: https://www.youtube.com/watch?v=0w0pi4-FRDY [<<< A bit bloody]

ratsbeyfus

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

skylla

ratsbeyfus wrote:

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

Chris Packham would agree, you should have stuck to fruit flies (although they are still very difficult to manipulate though)

ratsbeyfus

skylla wrote:

ratsbeyfus wrote:

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

Chris Packham would agree, you should have stuck to fruit flies (although they are still very difficult to manipulate though)

I'd had my drosophofill of fruit flies in the lab after three years (fascinating though they were) - plus I liked getting out into the 'field'. Transposed these yearnings in adult life into cyclo-X and avoiding getting on my turbo at all costs.

skylla

ratsbeyfus wrote:

skylla wrote:

ratsbeyfus wrote:

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

Chris Packham would agree, you should have stuck to fruit flies (although they are still very difficult to manipulate though)

I'd had my drosophofill of fruit flies in the lab after three years (fascinating though they were) - plus I liked getting out into the 'field'. Transposed these yearnings in adult life into cyclo-X and avoiding getting on my turbo at all costs.

geek

ratsbeyfus

skylla wrote:

ratsbeyfus wrote:

skylla wrote:

ratsbeyfus wrote:

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

Chris Packham would agree, you should have stuck to fruit flies (although they are still very difficult to manipulate though)

I'd had my drosophofill of fruit flies in the lab after three years (fascinating though they were) - plus I liked getting out into the 'field'. Transposed these yearnings in adult life into cyclo-X and avoiding getting on my turbo at all costs.

geek

...
printf("Thanks.\n"); /* Geek == Compliment */
...

jawooga

ratsbeyfus wrote:

skylla wrote:

ratsbeyfus wrote:

skylla wrote:

ratsbeyfus wrote:

OK, I've got a 20 year old Biology degree so what I say goes.

Hmmm...let me see... OK, I've read this thread and have formulated these two conclusions:

A. things have moved pretty quickly in the world of gene-manipulation since I played around with fruit-flies;

B. doing a final year project on the territorial distribution of faeces by Meles meles (aka badgers) was not as useful as I first thought it would be.

As you were...

Chris Packham would agree, you should have stuck to fruit flies (although they are still very difficult to manipulate though)

I'd had my drosophofill of fruit flies in the lab after three years (fascinating though they were) - plus I liked getting out into the 'field'. Transposed these yearnings in adult life into cyclo-X and avoiding getting on my turbo at all costs.

geek

...
printf("Thanks.\n"); /* Geek == Compliment */
...

C ? Old school, I like it !

ratsbeyfus

I loved coding in C... unfortunately the first (and last) job I took as a software engineer was in C++ (Symbian/EPOC) which I hated. Is C still used much? Apologies for going way OT...

jawooga

Not sure tbh. I love it too but not coded C for a while.

In an effort to nudge back on topic, the Guardian Sports section was talking about gene therapy in their special on Saturday so i guess it is widely perceived to be the next generation.