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Transgenic Lines Proven Unstable

press-release

 

The Institute of Science in Society

Science Society Sustainability

http://www.i-sis.org.uk

 

General Enquiries sam

Website/Mailing List press-release

ISIS Director m.w.ho

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Transgenic Lines Proven Unstable

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The insert in every commercially approved GM line has undergone rearrangement.

The cauliflower mosaic virus promoter plays a major role. This should be the

final nail in the coffin for GM crops, says Dr. Mae-Wan Ho

(m.w.ho), who has, for years, challenged scientific committees

advising governments over this very issue.

 

There is plenty of evidence that transgenic lines are unstable, which is why

ISIS has long recommended that appropriate molecular methods must be used to

document the stability of the GM insert before any transgenic line is released

into the environment. The characterization of the insert must be

‘event-specific’, which not only gives the structure of the insert, but also the

host genome sequences flanking the insert, proving that the insert remains

stable in successive generations. This recommendation has been incorporated into

the current European Directive (2001/18 /EC) on deliberate release of GMOs.

 

But to this day, pro-GM scientists advising the UK and other governments have

refused to acknowledge the evidence on transgenic instability, and worse. In its

latest reply to ISIS, the UK Advisory Committee on Releases to the Environment

(ACRE) has gone as far as to say that event-specific molecular characterization

is not necessary, thus going against the European Directive (see ISIS’ final

response to ACRE: Let the people decide http://www.i-sis.org.uk/ISISFRTA.php).

 

ISIS has reiterated 5 experiments which should be done to address the ‘areas of

uncertainty’, one of which calls for full event-specific molecular

characterization of all transgenic lines to establish uniformity and genetic

stability of the transgenic DNA insert(s), and " comparison with the original

data supplied by the biotech company to gain approval for field trials or for

commercial release. "

 

I am pleased to report that some effort has recently been made to do such

experiments by French scientists from the Laboratory of Methods for Detecting

GMOs in Versaille, and the Laboratory of Biometry and Artificial Intelligence,

Domaine de Vilvert in Jouy-en-Josas. And they have presented their results in a

poster at a conference in June 2003 [1].

 

The scientists recognized that, as labeling laws and thresholds are established

for foods containing GMOs in Europe, Japan, Australia, New Zealand and

elsewhere, " reliable GMO identification and quantification methods are needed to

comply with the regulations. " And " in order for these tests to be specific, the

sequence and detailed characterization of the GMO inserts and their edges are

required. "

 

Five different commercially approved GMOs in Europe were analyzed: three from

Monsanto, one from Bayer and one from Syngenta. All inserts were rearranged from

their intended gene order. Moreover, all five inserts showed further

rearrangements from the original structure submitted by the companies. In other

words, either the companies were mistaken about the original structure, or more

likely, further rearrangements had occurred after the crops had been

commercially grown. The details are given in Box 1.

 

 

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Box 1

 

Scrambling and further scrambling of GM inserts

T25 maize LibertyLink (Bayer)

 

Modified for tolerance to herbicide glufosinate. Company data showed insert

includes a truncated ampicillin resistance bla gene in the plasmid vector pUC18,

a CaMV 35S promoter (hereafter referred to as P35S) driving a synthetic pat gene

(glufosinate tolerance) terminated by CaMV 35S terminator (hereafter referred to

as T-35S). On analysis, the insert was found to have undergone further

rearrangement, so that a second, truncated and rearranged P35S has been joined

to the 5’ (left, or head) end of the insert, while additional pUC18 sequences

were found at the 3’ (right, or tail) end.

 

Edges flanking the insert show homologies (similarities) with Huck

retrotransposons (a class of mobile genetic elements) in the maize genome.

 

Mon 810 maize YieldGard (Monsanto)

 

Modified for resistance to lepidopteran insects (butterflies & moths). Company

data showed insert has a P35S driving a CrylAb synthetic gene with terminator

T-nos. Analysis revealed however, that T-nos and part of the 3’ (tail) end of

the CrylAb gene have been deleted. T-nos has been detected elsewhere in the

genome, indicating that it has moved from its original position.

 

The 5’ (head) end of the insertion site shows homology to the long terminal

repeats (LTR) of the maize alpha Zein gene cluster, but no homology to the maize

genome was detected at the 3’ site, indicating that there has been scrambling of

the maize genome at the insertion site.

 

GTS 40-3-2 soybean (Monsanto)

 

Modified for tolerance to herbicide glyphosate (Roundup Ready). Company data

showed insert with P35S driving a composite gene containing the N-terminal

chloroplast transit peptide (CPT4) joined to modified epsps gene with T-nos

terminator.Analysis revealed that a 254bp piece of DNA homologous to the epsps

gene and 534bp of unknown DNA have been joined to the 3’end of the insert.

 

It was not possible to identify the insertion site at all, indicating

substantial genome scrambling or deletion at the insertion site.

 

Bt 176 maize (Syngenta)

 

Modified for tolerance to herbicide glufosinate, male sterility and insect

resistance. The structures of two inserts, originating from two GM constructs,

were provided by the company. Only the simpler construct was analyzed. Company

data showed insert contains P35S driving the bar gene (glufosinate tolerance)

terminated by T35S, followed by the ampicillin resistance (bla) gene plus

bacterial promoter, and plasmid origin of replication, ori. Analysis revealed

several fragments, all containing CaMV 35S promoter, one with P35S joined to

T35S, a second with P35S joined to an unknown sequence, and a third with P35S

joined to the bar gene with the T35S deleted.

 

There were at least three insertion sites.

 

GA 21 maize (Monsanto)

 

Modified for tolerance to herbicide glyphosate (Roundup Ready). Company data

indicated insert contains multiple copies of the cassette with the rice actin

gene promoter (P-ract) driving the composite gene containing the N-terminal

chloroplast transit peptide (CPT4) joined to modified epsps gene and T-nos.

There were three complete cassettes flanked by a cassette with P-ract partially

deleted at the 5’ end, and one cassette with 3’ deletion of epsps plus a lone

P-ract at the 3’end. Analysis found partial deletion of P-ract and deletion of

T-nos in two different cassettes.

 

The insertion site at the 3’end is flanked by sequences of pol polyprotein gene

belonging to a PREM2-retrotransposon.

 

***********************************************************

 

The results revealed that,

 

All GMO inserts had rearranged from the structure provided by the company.

Many of the breakpoints for rearrangement involve the CaMV 35S promoter, as can

be predicted from its known recombination hotspot.

Scrambling of the genome at the site of insertion occurred in at least two out

of five inserts.

GMO inserts appear to show a preference for mobile genetic elements

(retrotransposons), with Long Terminal Repeats containing strong promoters,

which would result in " altered spatial and temporal expression patterns of

genes " nearby. In addition, it increases the chances that the inserts will move

with the retrotransposons, resulting in further genome scrambling and horizontal

gene transfer.

With considerable irony, whether intended or not is unclear, the authors

conclude: " Studying GMO’s structure is necessary to develop reliable

quantification and detection tests complying with the different regulations, but

it also leads [one] to ask fundamental questions about genome fluidity. Many of

the mechanisms involved in recombinant DNA integration are similar to those

underlying genome evolution. Therefore, characterized GMO inserts are a very

good model to study the molecular system involved in DNA rearrangements in

general. "

 

Collonier C, Berthier G, Boyer F, Duplan M-N, Fernandez S, Kebdani N, Kobilinsky

A, Romanuk M, Bertheau Y. Characterization of commercial GMO inserts: a source

of useful material to study genome fluidity. Poster courtesy of Pr. Gilles-Eric

Seralini, Président du Conseil Scientifique du CRII-GEN, www.crii-gen.org

 

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This article can be found on the I-SIS website at

http://www.i-sis.org.uk/TLPU.php

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General Enquiries sam

Website/Mailing List press-release

ISIS Director m.w.ho

 

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