Laws Governing Synthetic Biology

UK and EU Law

Synthetic biology has potential to help solve diverse problems and make the world a better place. This has been reflected by the eagerness of governments across the world to support and fund synthetic biology projects and initiatives.

However, the effects of genetic engineering are complex and currently difficult to predict. There is a risk that modified genes pass from our engineered organisms into wild strains, and out of our control; in plants through interbreeding, in bacteria through horizontal gene transfer. It has been mooted that genetically modified organisms may outcompete native species, reducing biodiversity, or that eating genetically modified crops may have long term adverse health effects, although there is currently little evidence for this.

The above concerns, coupled with worries from the public ranging from the ‘unnaturalness’ of genetic engineering to valid ethical and economic worries (as covered in our ethics section) have led to stringent regulations on the cultivation and release, whether controlled or into the environment, of genetically modified organisms.

In most countries, research groups/universities have committees for the approval of experiments using or producing genetically modified organisms. Higher risk experiments usually have to be ratified by a national regulatory body. In the UK, this is DEFRA, and in the U.S. the Coordinated Framework for the Regulation of Biotechnology. European assessments for approval are solely concerned with safety, whilst U.S. also considers the potential economic impact of the proposed experiments. The 6 steps in environmental risk assessment that must be followed before experimentation can take place in Europe are:

Identification of characteristics which may cause adverse effects
Evaluation of the potential consequences of each adverse effect, if it occurs
Evaluation of the likelihood of the occurrence of each identified potential adverse effect
Estimation of the risk posed by each identified characteristic of the GMO(s)
Application of management strategies for risks from the deliberate release or marketing of GMO(s)
Determination of the overall risk of the GMO(s)

It should be noted that there are various exempt dealings for laboratory based experiments on highly studied genetically modified organisms. The vast majority of iGEM projects will fit within this category- our project for one did not need to be certified by DEFRA.

Experiments with the aim of releasing the modified organisms are treated differently to those which merely aim to use the organism within a contained system. Note that this need not necessarily be in a research lab, but could be in a sealed vial.

European laws regarding the release of genetically modified organisms are notably stringent, with all organisms subject to extensive case by case evaluations by the European Food Safety Authority, and then passed through a number of other EU committees before being authorized for use.

Definition of GMOs in UK legislation

Techniques constituting genetic modification:

recombinant nucleic acid techniques involving the formation of new combinations of genetic material by the insertion of nucleic acid molecules, produced by whatever means outside an organism, into any virus, bacterial plasmid or other vector system and their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation;
techniques involving the direct introduction into an organism of heritable genetic material prepared outside the organism, including micro-injection, macro-injection and micro-encapsulation;
cell fusion or hybridization techniques where live cells with new combinations of heritable genetic material are formed through the fusion of two or more cells by means of methods that do not occur naturally.

Techniques which are not considered to result in genetic modification:

in vitro fertilisation;
natural processes including conjugation, transduction or transformation;
polyploidy induction.

Definition of GMOs in EU legislation

GMOs not being released into the environment

According to article 2(b) of EU Directive 2009/41/EC on the contained use of genetically modified micro-organisms:

‘Genetically modified micro-organism’ (GMM) means a micro-organism in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination; within the terms of this definition:

genetic modification occurs at least through the use of the techniques listed in Annex I, Part A;
the techniques listed in Annex I, Part B, are not considered to result in genetic modification;

Annex 1-Part A

Techniques of genetic modification:

Recombinant nucleic acid techniques involving the formation of new combinations of genetic material by the insertion of nucleic acid molecules produced by whatever means outside an organism, into any virus, bacterial plasmid or other vector system and their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation.
Techniques involving the direct introduction into a micro-organism of heritable material prepared outside the micro-organism, including micro-injection, macro-injection and micro-encapsulation.
Cell fusion or hybridisation techniques where live cells with new combinations of heritable genetic material are formed through the fusion of two or more cells by means of methods that do not occur naturally.

Annex 1-Part B

Techniques which are not considered to result in genetic modification:

in vitro fertilisation
natural processes such as: conjugation, transduction, transformation
polyploidy induction

GMOs being released into the environment

According to Article 2(2) of EU Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms:

“Genetically modified organism (GMO)” means an organism, with the exception of human beings, in which the genetic material has been altered in a way that does not occur naturally by mating and/or natural recombination.

Annex I A - Part 1

Techniques defined as genetic modification:

recombinant nucleic acid techniques involving the formation of new combinations of genetic material by the insertion of nucleic acid molecules produced by whatever means outside an organism, into any virus, bacterial plasmid or other vector system and their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation;
techniques involving the direct introduction into an organism of heritable material prepared outside the organism including micro-injection, macro-injection and micro-encapsulation;
cell fusion (including protoplast fusion) or hybridisation techniques where live cells with new combinations of heritable genetic material are formed through the fusion of two or more cells by means of methods that do not occur naturally.

Annex I A - Part 2

Techniques which are not considered to result in genetic modification:

in vitro fertilisation,
natural processes such as: conjugation, transduction, transformation,
polyploidy induction

US Law

The U.S.A is generally less risk averse to GMOs than European governments. U.S. Law is more relaxed regarding the risks associated with GMOs than European regulation. The law appears to mirror public perception across the two regions.

In the USA the National institute for health (NIH), an agency of the U.S. Department of Health & Human Services, has created guidelines for research involving recombinant or synthetic nucleic acid molecules. These govern all experiments in institutions receiving NIH support for recombinant or synthetic nucleic acid research or research being conducted on humans with recombinant or synthetic nucleic acid. According to section I-A of the guidelines:

‘The purpose of the NIH Guidelines is to specify the practices for constructing and handling: (i) recombinant nucleic acid molecules, (ii) synthetic nucleic acid molecules, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, and (iii) cells, organisms, and viruses containing such molecules.’

According to section I-B these guidelines, recombinant and synthetic nucleic acid molecules are defined as:

(i) molecules that a) are constructed by joining nucleic acid molecules and b) that can replicate in a living cell, i.e., recombinant nucleic acids;

(ii) nucleic acid molecules that are chemically or by other means synthesized or amplified, including those

that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid

molecules, i.e., synthetic nucleic acids, or

(iii) molecules that result from the replication of those described in (i) or (ii) above

Interestingly, by this definition it would appear that the genomes of organisms that had undergone genome shuffling may not be classified as recombinant or synthetic molecules of DNA. This would mean they are not covered by these guidelines set out by the NIH.

Of course the genomes that had been shuffled should not have been the subject of genetic engineering themselves.

Whilst genome shuffling is not directly addressed in U.S. law, researchers in the area claim that it does not count as genetic engineering, and state this as a major an advantage e.g. Xang et al.

The Presidential Commission on Bioethics has released a report into synthetic biology. This called for greater federal oversight of this new field. The commission also called for greater public education about synthetic biology and greater ethical education for individuals in the field.

Implications for BareCillus

Whilst the L-forms we have created with our ‘switch’ BioBrick could not currently be released into the environment under EU and UK laws- extensive testing would need to be done and our BioBrick contains an antibiotic resistance marker (which cannot be released into the environment)- L-forms of many species occur naturally. These bacteria are ‘natural’ L-forms.

Part of our project involves the fusing of L-forms to promote genome shuffling and directed evolution. Would the fusion of two natural L-forms constitute genetic engineering? In theory, fusion could occur very rarely naturally. Arguably our project is very similar to fertilization, with two cells fusing and similar genomes mixing. It is also similar to meiosis, with genome shuffling nearly indistinguishable from crossing over. In which case, could L-forms that have undergone rounds of genome shuffling count as genetically engineered?

UK law appears clear on this. The following is cited as an example of genetic modification:

cell fusion or hybridization techniques where live cells with new combinations of heritable genetic material are formed through the fusion of two or more cells by means of methods that do not occur naturally.

However are the methods by which we are inducing fusion unnatural? On one hand, we are physically pushing the L-forms together, and PEG is required to put shear stress of the cell membranes. But fusion can occur naturally in L-forms. Furthermore whilst the legislation is fairly clear, there have been no cases yet, and so it is not fully clear how the laws should be interpreted, or whether the legislation has any wiggle room or could be challenged.

Cells that have undergone genomic fusion are exempt from many of the regulations that other genetically modified organisms are subject to, as stated in ‘The Genetically Modified Organisms (Contained Use) Regulations 2000’. This would be useful for researchers hoping to use L-forms for directed evolution. It should be noted however that random mutagenesis is also exempt from all but regulation 17, as can be found here.