Biological diversity, including marine biological diversity, is essential for human life. However, presently biological diversity on land and in the oceans is rapidly declining. Thus, there is a strong need to establish legal frameworks for the conservation of marine biological diversity. In this regard, growing attention is being paid to the establishment of marine protected areas (MPAs). This chapter will explore emergent norms on this subject. In particular, the following issues will be examined: (1) the principal approaches to conservation of marine biological diversity, (2) the limits of the LOSC with regard to the conservation of marine biological diversity, (3) the significance of, and the limitations associated with, the Convention on Biological Diversity in the context of the conservation of marine biological diversity, (4) MPAs and their limitations, and (5) MPAs on the high seas.

Chapter 14 describes the biotechnological applications of recombinant DNA technology. The range of disciplines that contribute to biotechnology is outlined to illustrate the scale and scope of the sector. Production of proteins is one key area where cloned genes can be expressed to produce high-value products for use in a variety of applications, and the types of systems used for protein production are discussed. Protein engineering by methods such as rational design and directed evolution has enabled customised proteins to be developed for specific applications. The requirements for transition from laboratory-scale research and development to industrial production at a commercially viable level are outlined, and the contribution of the biotechnology sector in managing the COVID-19 pandemic is discussed.

In Chapter 8, various strategies that can be used to clone DNA fragments are described. Cloning genomic DNA and complementary DNA (cDNA) to generate libraries of cloned fragments remain two of the most common methods for primary library construction. Fragments may also be generated by polymerase chain reaction (PCR), or may be designed from a sequence database and synthesised in vitro. The choice of vector (plasmid, bacteriophage, virus or artificial chromosome) depends on the intended outcome, the size and origin of fragments and whether it is a primary cloning or a sub-cloning protocol. Restriction-dependent and restriction-independent methods can be used to join fragments to vectors. Techniques such as Golden Gate cloning, Gateway technology and Gibson assembly have mostly replaced earlier methods and can be used to assemble several fragments into a multi-fragment construct.

In Chapter 17, the topic of organismal cloning is described, noting the difference between reproductive and therapeutic cloning. Preformationism and epigenesis (as concepts of development), and the work in the early 1900s that led to the development of the concept of nuclear totipotency, are outlined. As the concept was refined, cells that are pluripotent, multipotent or irreversibly differentiated were described. Cloning using nuclear transfer was first proposed in 1938, and achieved in 1952. More correctly known as somatic cell nuclear transfer (SCNT), the birth of Dolly in 1996 was a milestone in that she was the first mammal to be cloned using a fully differentiated somatic cell as the source of the donor nucleus.

Chapter 3 outlines the need for a broad ethical framework in all areas of human activity, in particular the importance of ethics and bioethics in the context of genetic engineering. The basis of the ethical framework is outlined, and some key ethical issues across the eras of genetics are described.Current and emerging ethical dilemmas, and the need for active engagement with ethical issues in science, by both scientists and the public, are noted.

In Chapter 7, the host cells and vectors that are used to enable recombinant DNA to be propagated and amplified are discussed. A range of prokaryotic and eukaryotic cells can be used to propagate DNA in vectors derived from plasmids, bacteriophages and plant and animal viruses. Vectors are engineered to have particular properties such as unique cloning sites, origins of replication, selectable markers and promoters for expressing cloned genes. Vectors are designed to be compatible with one or more host cell types to enable flexibility in use. When recombinant DNA has been generated in vitro, the vector/insert combination is introduced into the target host cell by processes such as transformation and transfection. Alternatively, a mechanical process can be used, such as biolistic delivery using a ‘gene gun’ to fire DNA directly into cells.

In Chapter 10, the methods used to identify and select specific clones from clone banks (libraries) are described. The terms selection and screening are defined, and selection using selectable markers for antibiotic resistance is outlined as an example. Other genetic selection methods such as use of chromogenic substrates, insertional inactivation and complementation of defined mutations are described. Screening clone banks with nucleic acid probes and hybridisation methods is outlined to illustrate the powerful and specific nature of this approach. Radioactive and non-radioactive labelling techniques are compared, with the advantages and disadvantages of each outlined. The use of the polymerase chain reaction in screening, and immunological screening for expressed genes are described. Automation of screening is discussed to illustrate how high-throughput screening protocols enable very large numbers of clones to be screened efficiently. Analysis of cloned genes is described, covering techniques such as restriction mapping, Southern blotting and its derivative methods, sub-cloning and DNA sequencing.

The generation of transgenic plants and animals is discussed in Chapter 16. The technology is now well established, but remains a controversial area in terms of public perception and acceptance. Scientific, regulatory, ethical, political and commercial factors together present a complex framework within which the development of transgenic plants and animals is placed. As well as considering the technical procedures used to generate transgenics, these broader aspects (and their impact on the success or failure of a transgenic product) are considered. The development of Golden Rice, and the subsequent political issues around its deployment, illustrate the complexity of the topic. Paradoxically, transgenic animals often generate less negative reaction from the anti-GMO (genetically modified organism) movement than transgenic plants, with most concerns being around animal welfare issues. This is particularly true where the product is, for example, a demonstrably positive therapeutic; when transgenic animals are generated for consumption, the GMO debate tends to become polarised to the same extent as transgenic crops.

This chapter will examine rules governing marine spaces beyond the limits of national jurisdiction, namely, the high seas and the Area. The high seas are essentially characterised by the principle of freedom of the seas, and order in the high seas is ensured primarily by the flag State. Thus, the principle of the exclusive jurisdiction of the flag State and its exceptions are key issues underlying international law governing the high seas. However, the Area is governed by the principle of the common heritage of mankind. This principle is innovative because it may bring new viewpoints beyond the State-to-State perspective in the law of the sea. Against that background, this chapter will discuss in particular the following issues: (1) the principle of freedom of the high seas, (2) the principle of the exclusive jurisdiction of the high seas, (3) the problems associated with flags of convenience, (4) the peacetime exceptions to the principle of the exclusive jurisdiction of the flag State on the high seas, (5) the raison dtre of the principle of the common heritage of mankind, and (6) the 1994 Implementation Agreement

Peaceful settlement of international disputes occupies an important place in international law in general and the law of the sea is no exception. In this respect, the LOSC establishes a unique mechanism combining the voluntary and compulsory procedures for dispute settlement. It is particularly significant that the LOSC sets out the compulsory dispute settlement procedures as an integrated part of the Convention. Furthermore, it is of particular interest to note that a new permanent international tribunal, namely, ITLOS, was established. The dispute settlement procedures of the LOSC provide an interesting insight into the development of dispute settlement procedures in international law. Thus, this chapter will address the dispute settlement procedures under the LOSC with particular reference to the following issues: (1) the principal features of the dispute settlement procedures of the LOSC, (2) the significance of and limitations associated with the compulsory procedures for dispute settlement under the LOSC, and (3) jurisdiction over mixed disputes involving maritime and non-maritime issues, and (40 a risk of fragmentation of international law.

Marine living resources are of vital importance for mankind because these resources are an essential source of protein and many human communities depend on fishing. As marine living resources are renewable, there is certainly a need to pursue conservation policies in order to secure sustainable use of these resources. Nonetheless, the depletion of these resources is a matter of more pressing concern in the international community. Thus, the conservation of marine living resources is a significant issue in the law of the sea. This chapter will examine rules of international law governing the conservation of these resources, focusing particularly on the following issues: (1) the problems associated with the traditional approaches to conservation of marine living resources, (2) the concept of sustainable development living resources, (3) the ecosystem approach, (4) the precautionary approach, and (5) the role of regional fisheries management organisations in the conservation of marine living resources and its main challenges.

In Chapter 12, the techniques used for editing genomes are discussed, from early gene targeting approaches to highly specific CRISPR and prime editing techniques using engineered nucleases. The development of techniques through zinc-finger nucleases and TALENs is used to set the context for the CRISPR and prime editing methods that have become widely accessible and are used extensively to make changes to genomes. As an alternative to editing the genome, RNA editing offers a different approach that enables modulation of gene expression without complete inactivation or alteration of genes and may be more appropriate in some therapeutic situations. The potential for genome and RNA editing is outlined, and the ethical aspects noted.