Developments in genetic modification of cattle and implications for regulation, safety and traceability

doi:10.15302/J-FASE-2019306

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Abstract：

Genetic modification techniques, in particular novel gene editing technologies, hold the yet unfulfilled promise of altering genetic traits in farm animals more efficiently than by crossbreeding, allowing for a more rapid development of new cattle breeds with distinct traits. Gene editing technologies allow for the directed alteration of specific traits and thereby have the potential to enhance, for instance, disease resilience, production yield and the production of desired substances in milk. The potential implications of these technological advancements, which are often combined with animal cloning methods, are discussed both for animal health and for consumer safety, also with consideration of available methods for the detection and identification of the related products in the food supply chain. Finally, an overview is provided of current regulatory approaches in the European Union (EU) and major countries exporting beef to the EU, for products from animals bred through established practices as well as modern biotechnologies.

Keywords cattle food safety gene editing genetic modification GMO detection regulation

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	Jan Pieter VAN DER BERG
	Gijs A. KLETER
	Evy BATTAGLIA
	Martien A. M. GROENEN
	Esther J. KOK

引用本文:

Jan Pieter VAN DER BERG,Gijs A. KLETER,Evy BATTAGLIA, et al. Developments in genetic modification of cattle and implications for regulation, safety and traceability[J]. Front. Agr. Sci. Eng. , 2020, 7(2): 136-147.

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https://journal.hep.com.cn/fase/EN/10.15302/J-FASE-2019306 OR https://journal.hep.com.cn/fase/EN/Y2020/V7/I2/136

Trait	Method	Trait	Reference
Milk composition and human	Transgenesis using microinjection	Introduction of gene encoding human lactoferrin	^[¹⁵^]
Protein production	Transgenesis using microinjection	Introduction of gene encoding human α-lactalbumin	^[¹⁶^,¹⁷^]
	Transgenesis in somatic cells, SCNT	Introduction of gene encoding human bile salt-stimulated lipase	^[¹⁸^]
	Transgenesis in somatic cells, SCNT	Introduction of gene encoding human immunoglobulin	^[¹⁹^,²⁰^]
	Transgenesis in somatic cells, SCNT	Introduction of additional gene copies encoding bovine α- & k-casein	^[²¹^]
	Transgenesis in somatic cells, SCNT	Introduction of gene encoding human lysozyme	^[²²^]
	Gene editing using zinc finger nucleases, NHEJ repair, SCNT	Disruption of β-lactoglobulin gene	^[¹²^]
	Transgenesis in somatic cells, SCNT	Introduction of gene encoding humanized Caenorhabditis elegans n-3 fatty acid desaturase	^[¹¹^]
	Transgenesis in somatic cells, SCNT	Introduction of gene encoding human β-defensin-3	^[²³^]
	Transgenesis in somatic cells using TALENs & SCNT	Introduction of gene encoding Sulfolobus solfataricus lactase	^[²⁴^]
Disease resilience
- Mastitis	Transgenesis in somatic cells, SCNT	Introduction of gene encoding Staphylococcus simulans lysostaphin	^[²⁵^]
- Bovine spongiform encephalopathy	Transgenesis and embryonic cloning	Disruption of prion protein via integration of knockout vectors	^[²⁶^]
- Mannheimia hemolytica leukotoxin	Gene editing and homology-directed repair, SCNT	Gene-edited CD18, substitution of a glutamine for a glycine codon in its signal peptide	^[²⁷^]
- Bovine tuberculosis	Transgenesis in somatic cells using TALENs & SCNT	Introduction of mouse nuclear body protein encoding gene SP110	^[²⁸^]
	Transgenesis in somatic cells using Cas9n & SCNT	Introduction of additional genes encoding solute carrier family gene NRAMP1	^[¹³^]
Hornlessness	Gene editing and homology-directed repair, SCNT	Introduction of bovine P_c POLLED allele, resulting in hornless phenotype	^[²⁹^]
Thermotolerance	Gene editing using TALENs, SCNT	Introduction of the SLICK locus for improved thermotolerance	^[³⁰^,³¹^]
Increased muscle growth	Gene editing using TALEN mRNA	Introduction of small deletions in the myostatin (GDF8) gene by use of gene editing	^[¹⁴^]
Other fundamental research
-Reverse transcribed gene transfer	Transgenesis via pronuclear injection of retroviral vector DNA	Integration of hepatitis B surface antigen gene using a retroviral vector	^[³²^]
-Marker assisted selection	Transgenesis via transfection of somatic cells with retroviral vector DNA, SCNT	Integration of a β-galactosidase-neomycin fusion gene driven by cytomegalovirus promoter	^[³³^]
-Lentiviral infection	Transgenesis in somatic cells using lentiviral vectors, SCNT	Integration of eGFP using lentiviral vector	^[³⁴^]
-Transposon integration	Transgenesis via microinjection of transposon DNA	Sleeping Beauty and Piggybac transposons used to deliver sequences containing fluorescent protein genes	^[³⁵^]
-Targeted integration	Transgenesis in somatic cells using TALENs & SCNT	Integration of eGFP in Rosa26 “safe locus”	^[³⁶^]

Tab.1 Examples of transgenic and gene-edited cattle

Fig.1 Timeline of technological developments and milestones used to obtain GM cattle. MI, pronuclear microinjection; SCNT, somatic cell nuclear transfer; SDN, site-directed endonucleases. Important milestones are summarized on the right.

Country	Animal cloning	Transgenic livestock	Gene-edited livestock	Reference
EU member states	Prohibited, until specific regulations on animal cloning are in place	Requires approval according to EU Directive 2001/18/EC and Regulation (EC) No. 1829/2003, safety assessment performed by EFSA GMO Panel	Requires approval according to EU Directive 2001/18/EC and Regulation (EC) No. 1829/2003, safety assessment performed by EFSA GMO Panel	^[⁵²^]
USA	Allowed, a risk management plan and guidance for industry have been issued by the FDA	Requires approval according to Federal FD&C Act, regulations for new animal drugs as stated in 2009 FDA Guidance for industry #187 (Draft guidance) and NEPA	Requires approval according to Federal FD&C Act, regulations for new animal drugs as stated in 2009 FDA Guidance for industry #187 (Draft guidance) and NEPA	^[⁵³^–⁵⁵^]
Canada	Allowed, food products of cloned animals and clone progeny are considered “novel foods” and require pre-market safety assessments according to the regulations in Division 28, Part B, of the Food and Drug Regulations (Novel Foods)	Requires approval according to the Canadian Environmental Protection Act, 1999, the New Substances Notification Regulations (Organisms) and Food and Drugs Act	No specific policy on gene editing, may be considered “novel” and require case-by-case safety assessment by Health Canada	^[⁵⁶^,⁵⁷^]
Argentina	Allowed	Requires approval according to animal biotechnology regulation, case-by-case assessment by CONABIA	Requires approval according to animal biotechnology regulation, case-by-case assessment by CONABIA	^[⁵⁸^]
Brazil	Allowed, commercial animal cloning mostly in partnership with EMBRAPA, registration of cloned cattle at ABCZ	Requires approval according to animal biotechnology regulation, case-by-case assessment by CTNBio	Requires approval according to animal biotechnology regulation, case-by-case assessment by CTNBio, gene-edited animals lacking recombinant DNA are regarded non-GM according to Normative Resolution #16	^[⁴⁶^]
Australia & New Zealand	Allowed, generally in confined research environment	Requires approval according to Gene Technology Act 2000, by OGTR	Requires approval according to Gene Technology Act 2000, by OGTR, gene editing techniques that do not introduce new genetic material are not regulated as GMOs	^[⁵⁹^,⁶⁰^]
Uruguay	No specific legislation on animal cloning, animal biotechnology performed in research institutes such as Institut Pasteur in Montevideo and the Animal Reproduction Institute of Uruguay	No specific legislation on animal biotechnology, environmental release of GMOs and biosecurity is subject to prior authorization by competent authorities, as stated in article 23 of law No. 17283 on the protection of the environment	No specific legislation on gene editing in animals, during a meeting of the CAS the minister of agriculture signed a declaration in favor of gene editing. Gene-edited animals may be subject to prior authorization according to law No. 17283	^[⁶¹^,⁶²^]

Tab.2 Regulation of animal cloning, transgenesis and gene editing

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