Send to Environmental Protection Authority preferably by email (neworganisms@epa.govt.nz) or alternatively by post (Private Bag 63002, Wellington 6140)
Payment must accompany final application; see our fees and charges schedule for details.
To obtain approval for projects to develop
genetically modified organisms in containment
Application Number
Date
Completing this application form
1. This form has been approved under section 42A of the Hazardous Substances and New
Organisms (HSNO) Act 1996. It only covers projects for development (production, fermentation or regeneration) of genetically modified organisms in containment. This application form may be used to seek approvals for a range of new organisms, if the organisms are part of a defined project and meet the criteria for low risk modifications. Low risk genetic modification is defined in the HSNO (Low Risk Genetic Modification) Regulations:
http://www.legislation.govt.nz/regulation/public/2003/0152/latest/DLM195215.html.
2. If you wish to make an application for another type of approval or for another use (such as an emergency, special emergency or release), a different form will have to be used. All forms are available on our website.
3. It is recommended that you contact an Advisor at the Environmental Protection Authority (EPA) as early in the application process as possible. An Advisor can assist you with any questions you have during the preparation of your application.
4. Unless otherwise indicated, all sections of this form must be completed for the application to be formally received and assessed. If a section is not relevant to your application, please provide a comprehensive explanation why this does not apply. If you choose not to provide the specific information, you will need to apply for a waiver under section 59(3)(a)(ii) of the HSNO Act. This can be done by completing the section on the last page of this form.
5. Any extra material that does not fit in the application form must be clearly labelled, cross- referenced, and included with the application form when it is submitted.
6. Please add extra rows/tables where needed.
7. You must sign the final form (the EPA will accept electronically signed forms) and pay the application fee (including GST) unless you are already an approved EPA customer. To be recognised by the EPA as an “approved customer”, you must have submitted more than one application per month over the preceding six months, and have no history of delay in making payments, at the time of presenting an application.
8. Information about application fees is available on the EPA website.
9. All application communications from the EPA will be provided electronically, unless you specifically request otherwise.
Commercially sensitive information
10. Commercially sensitive information must be included in an appendix to this form and be identified as confidential. If you consider any information to be commercially sensitive, please show this in the relevant section of this form and cross reference to where that information is located in the confidential appendix.
11. Any information you supply to the EPA prior to formal lodgement of your application will not be publicly released. Following formal lodgement of your application any information in the body of this application form and any non-confidential appendices will become publicly available.
12. Once you have formally lodged your application with the EPA, any information you have supplied to the EPA about your application is subject to the Official Information Act 1982 (OIA). If a request is made for the release of information that you consider to be confidential, your view will be considered in a manner consistent with the OIA and with section 57 of the HSNO Act. You may be required to provide further justification for your claim of confidentiality.
Definitions
Containment
Restricting an organism or substance to a secure location or facility to prevent escape. In respect to genetically modified organisms, this includes field testing and large scale fermentation
Controls
Any obligation or restrictions imposed on any new organism, or any person in relation to any new organism, by the HSNO Act or any other Act or any regulations, rules, codes, or other documents made in accordance with the provisions of the HSNO Act or any other Act for the purposes of controlling the adverse effects of that organism on people or the environment
Genetically Modified Organism (GMO)
Any organism in which any of the genes or other genetic material:
• Have been modified by in vitro techniques, or
• Are inherited or otherwise derived, through any number of replications, from any genes or other genetic material which has been modified by in vitro techniques
New Organism
A new organism is an organism that is any of the following:
• An organism belonging to a species that was not present in New Zealand immediately before 29 July 1998;
• An organism belonging to a species, subspecies, infrasubspecies, variety, strain, or cultivar prescribed as a risk species, where that organism was not present in New Zealand at the time of promulgation of the relevant
regulation;
• An organism for which a containment approval has been given under the HSNO Act;
• An organism for which a conditional release approval has been given under the HSNO Act;
• A qualifying organism approved for release with controls under the HSNO Act;
• A genetically modified organism;
• An organism belonging to a species, subspecies, infrasubspecies, variety, strain, or cultivar that has been eradicated from New Zealand;
• An organism present in New Zealand before 29 July 1998 in contravention of the Animals Act 1967 or the Plants Act 1970. This does not apply to the organism known as rabbit haemorrhagic disease virus, or rabbit calicivirus A new organism does not cease to be a new organism because:
• It is subject to a conditional release approval; or
• It is a qualifying organism approved for release with controls; or
• It is an incidentally imported new organism
Project An individual or collaborative endeavour that is planned to achieve a particular aim or research goal
1. Applicant details
1.1. Applicant
Company Name: (if applicable) Victoria University of Wellington Contact Name: Peter Pfeffer
Job Title: Associate Professor
Physical Address: School of Biological Sciences, Kelburn Parade, Wellington Postal Address (provide only if not the same as the physical):
Phone (office and/or mobile): 0224231054 Fax:
Email: peter.pfeffer@vuw.ac.nz
1.2. New Zealand agent or consultant (if applicable)
Company Name:
Contact Name:
Job Title:
Physical Address:
Postal Address (provide only if not the same as the physical): Phone (office and/or mobile):
Fax:
Email:
2. Information about the application
2.1. Brief application description
Use of recombinant lentiviral vectors as an efficient and lineage specific means to genetically modify cells and embryos for research applications related to mammalian development and biotechnological improvements for generating modified livestock.
2.2. Summary of application
This application covers the development and research use of replication-deficient
recombinant lentiviral vectors in containment. Vectors will be used as tools to genetically modify cells and embryos of various vertebrates in containment. Applications for
lentivirus include fundamental research to understand early mammalian development and applied research to investigate and improve animal production characteristics.
2.3. Technical description
Briefly describe the host organism(s) and the proposed genetic modifications. Please make sure that any technical words used are included in a glossary. Note if any part of this research project is already covered by an existing HSNO Act approval that your organisation holds or uses.
The overall purpose of the project is to utilize recombinant lentiviral vectors as a tool to genetically modify animal cells and embryos for research aiming to
A. Understand early vertebrate development (particularly with a long term view to increasing embryo survival in livestock with potential spinoff benefits for human pregnancy associated problems).
B. Generate animal models (mice, rats) for understanding the function of genes, to create animal models of human diseases and medical conditions and for commercial and medical applications (confer disease resistance, understand genes involved in and then enhance livestock traits, produce biopharmaceuticals, generate animal models of human diseases and for xenotransplantation).
There are four phases for this work:
1. Development of plasmids, including Lentiviral packaging and expression plasmids in Escherichia coli.
2. The characterisation of these plasmids by transfection of mammalian cell lines for analysis of gene expression and activity.
3. The packaging of recombinant Lentiviral vectors in mammalian cell lines.
4. The genetic modification of mammalian cell lines, embryos in containment using lentiviral vectors with genetically modified mouse and rat embryos being allowed to develop to term if required.
Background
Our research in livestock focuses on two areas A. The enhancement of livestock fertility B. Generation of animal models.
Area A involves basic long-term research where we are aiming to understand at a molecular (gene regulatory) and cellular level, causes of low pregnancy outcomes. It has been established that early embryo survival is one of the most critical factors involved in low fertility in most mammals. To this end we are presently conducting research that is pinpointing genes important to early embryogenesis and pregnancy establishment. In the next phase we need to understand the regulation of these genes and their in vivo function. For these purposes we need to genetically modify cells, embryos or whole animals so as to be able to monitor gene expression or perform loss and gain of function experiments.
Area B, the the generation of animal models is important for basic and translational research including the generation of transgenic mice and rats that mimic human disease states and susceptibilities (for example the creation of mice that are susceptible to SARS-CoV-2 by virtue of oexpressing the human ACE-2 receptor).
Why use lentiviral vectors?
1. Lentiviral vectors are one of the most efficient systems for introducing DNA into mammalian cells, including non-dividing cells and the third generation lentiviral system we propose to use is a routine molecular biological tool in use for over a decade (Dull et al., 1998; Park, 2007).
2. For studying early embryogenesis, infection at the blastocyst stage results in only the outer (trophoblast or placenta-forming) cells taking up DNA thus representing a unique opportunity in specifically targeting this tissue (Georgiades et al., 2007).
Lentivector particles are generated by co-expressing (i) the virion packaging elements and (ii) the vector genome in a cell used as producer, for instance a 293 or 293T human embryonic kidney cell. This results in transcriptional-replication of the vectorand packaging into infectious, lentiviral particles. The lentiviral supernatants produced by the transfected packaging cells can then be used to infect and transduce target cells Once the lentivirus enters the target cell, the recombinant viral- vector RNA is reverse-transcribed, actively imported into the nucleus and stably integrated into the host genome to express the DNA inserted into the vector. Due to various strategies, the vector in the lentiviral particles cannot replicate in the target cells.
The core and enzymatic components of the virion packaging elements come from HIV-1, while the envelope is derived from a heterologous virus, most often vesicular stomatitis virus (VSV) due to the high stability and broad host range of its G protein. For maximal biosafety reasons, gag, coding for the virion main structural proteins and pol, responsible for the retrovirus-specific enzymes are present on one plasmid, rev, which encodes a post-transcriptional regulator necessary for efficient gag and pol expression on a second plasmid and the envelope protein on a third. Although the three packaging plasmids allow expression in trans of proteins required to produce viral progeny (e.g. gal, pol, rev, env) in the 293 producer cell line, none of them contain
LTRs or the Ψ packaging sequence. This means that none of the HIV-1 structural genes are actually present in the packaged viral genome, and thus, are never expressed in the transduced target cell. No new replication-competent virus can be produced. The developed lentiviral vectors confirm to the conditions of the APP202444 decision for not being organisms.
The vector itself is the only genetic material transferred to the target cells. The transducing expression vector, into which the candidate gene coding sequence is inserted, contains modified HIV 5’ and 3’ LTRs, Rous sarcoma virus or CMV enhancer/ promoter for production of viral RNA in producer cells, HIV-1 Ψ packaging sequence, and antibiotic resistance genes for selection in E.
coli and mammalian cells. A deletion in the 3’LTR results in a self-inactivating virus.
Description of GMOs to be developed:
1) E. coli non-pathogenic laboratory strains (e.g., K12 and B non-conjugative strains) modified with Lentiviral packaging and vector plasmids.
2) Producer cells (e.g. HEK 293) transfected with Lentiviral packaging and vector plasmids to produce high titers of packaged lentiviral particles (= virions).
3) Target cells and embryos: mammalian primary cells and embryos in containment will be infected in vitro with lentiviral particles (“the vector”) resulting in integration of the vector LTRs and the sequences between these regions into the host genome. These sequences contain mammalian genetic material expressed during development or required for the realisation of our research goals outlined above. The vectors will also include DNA features commonly used in the art of genetic engineering such as promoters, reporter and selection marker genes, expression tags, secretory and targeting signals, recombination elements, and other gene regulatory elements from bacteria, fungi, viruses, invertebrates and vertebrates.
3. Information about the new organism(s)
3.1. Identity of the host organism(s) For each host organism:
• Provide its taxonomic name and describe what type of organism it is.
• Provide a description of the strain(s) being applied for (if relevant).
• If the host organism is derived from humans (eg, cell lines) or may have cultural significance (e.g.
sourced from native biota), provide details of its source.
• State the category (Category 1 or Category 2) of the host organism (as per the HSNO (Low Risk Genetic Modification) Regulations).
Latin binomial Escherichia coli (Migula 1895; Castellani and Chalmers, 1919).
Common name(s) E. coli
Type of organism Bacterium
Taxonomy Gamma Proteobacteria, Enterobacteriales, Enterobacteriaceae
Strain(s) Non-pathogenic laboratory strains (e.g., Genetically crippled derivatives of Escherichia coli K12 and strain B)
Other information There are no known inseparable or associated organisms.
HSNO Category 1
Latin binomial Mus musculus (Linnaeus, 1758)
Common name(s) Mouse
Type of organism Animal, embryos, animal cells
Taxonomy Mammalia, Rodentia, Muridae
Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from laboratory strains originally procured from reputable commercial sources. Mice are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos, adults: 2
Latin binomial Rattus rattus (Linnaeus, 1758)
Common name(s) Rat
Type of organism Animal, embryos, animal cells
Taxonomy Mammalia, Rodentia, Muridae
Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from laboratory strains originally procured from reputable commercial sources. Rats are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos, adults: 2
Latin binomial Homo sapiens (Linnaeus, 1758)
Common name(s) Human
Type of organism Animal cell
Taxonomy Mammalia, Primates, Hominidae
Strain(s) Commercially available cell lines (e.g., HEK293, HeLa)
Other information Human cell lines will be obtained from reputable commercial suppliers or research institutes. Only pure cell lines will be used for virus production and they do not contain any inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Oryctolagus cuniculis (Linnaues, 1758)
Common name(s) Rabbit
Type of organism Embryo, animal cell
Taxonomy Mammalia, Lagomorpha, Leporidae
Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from laboratory strains originally procured from reputable commercial sources. Rabbits are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Bos taurus (Linnaeus, 1758) Common name(s) Cattle, cow
Type of organism Embryo, animal cell
Taxonomy Mammalia, Artiodactyla, Bovidae Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from New Zealand cows or derived by in vitro fertilisation using commercially available bull semen with eggs sourced from abbatoirs or retrieved by transvaginal recovery. Cattle are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Capra hircus (Linnaeus, 1758) Common name(s) Domestic goat
Type of organism Embryo, animal cell
Taxonomy Mammalia, Artiodactyla, Bovidae Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from New Zealand goats or derived by in vitro fertilisation using commercially available goat semen with eggs sourced from euthanised animals or by transvaginal recovery. Goats are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Ovis aries (Linnaeus, 1758)
Common name(s) Sheep
Type of organism Embryo, animal cell
Taxonomy Mammalia, Artiodactyla, Bovidae Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from New Zealand sheep or derived by in vitro fertilisation using commercially available ram semen with eggs sourced from euthanised animals or by transvaginal recovery. Sheep are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Cervus elaphus (Linnaeus, 1758)
Common name(s) Red deer.
Type of organism Embryo, animal cell
Taxonomy Mammalia, artiodactyla, Cervidae Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes or derived in-house. Embryos will be retrieved from New Zealand deer or derived by in vitro fertilisation using commercially available deer semen with eggs sourced from euthanised animals or by transvaginal recovery. Deer are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
Latin binomial Gallus gallus (Linnaeus, 1758) Common name(s) Domestic chicken
Type of organism Embryo, animal cell
Taxonomy Aves, Galliformes, Phasianidae Strain(s)
Other information Cell lines will be obtained from reputable commercial suppliers or research institutes. Fertilised eggs will be sourced from commercial breeders and incubated in-house with embryos either kept in the egg or grown in culture for and after lentivirus transfection. Chickens are well characterised and do not contain inseparable or associated organisms.
HSNO Category 1, embryos: 2
3.2. Regulatory status of the organism
Is the organism that is the subject of this application also the subject of:
An innovative medicine application as defined in section 23A of the Medicines Act 1981?
☒
Yes☐
NoAn innovative agricultural compound application as defined in Part 6 of the Agricultural Compounds and Veterinary Medicines Act 1997?
☐
Yes☐
No4. Information about the project
4.1. Describe the nature and range of the proposed genetic modifications
• Describe the nature and range of the proposed genetic modifications (e.g. the range of elements that the vectors or gene constructs may contain, and the type, source and function of the donor genetic material).
• State the category (Category A or Category B) of the genetic modifications (as per the HSNO (Low Risk Genetic Modification) Regulations).
Vector system used, eg cloning or expression, plasmid, or viral
Escherichia coli will be developed using standard cloning and transformation techniques and will employ standard non-conjugative plasmid vectors from commercial and reputable research laboratory sources.
Non-replicative recombinant lentiviral vectors will be packaged using calcium phosphate-, liposome- or electroporation-mediated transfection of HEK293 cells with non-conjugative vector and packaging plasmid DNAs. The production of Lentiviral vectors using cell lines such as HEK293 and Lentiviral packaging plasmids is a Category B genetic modification as this modification increases the infectivity of the host as infectious particles are produced. Due to the genetic material exclusions, this research will fall under the Low Risk Regulations.
Cells and embryos contained in vitro will be genetically modified by transduction with high titer lentivirus particle containing supernatants from the above HEK-transfected cells. Transduction involves co- incubation of cells/embryos with lentiviral stock for 6 to 24 hours followed by a media change with appropriate safe disposal of infective lentivirus-containing medium. Category A.
Genetically modified embryos in utero and live animals (latter: mice and rats only) will be developed only by transferring in vitro transduced embryos into recipient animals. Embryo washings prior to transfer will ensure minimal exposure of recipient animal to infectious non- replicative lentivirus. Modification of chick embryos will also be performed in ovo (in the egg), but not allowed to develop beyond mid- gestation (10days). The vertebrate embryos used here and adult mice are Category 2 hosts as they are, or are able to grow into, a whole animal without human intervention. The genetic modification is thus a category B modification. Transfer of embryos previously modified with replication-defective lentivirus will not increase pathogenicity, infectivity (infectious particles will not be produced) or virulence of the host, will not result in a host with a greater ability to escape from containment, and the introduced sequences will be well characterised with respect to gene sequence and function thus meeting the requirements for low risk as stipulated under HASNO (SR 2003/152):
clauses 5(2)(b) AND 5(4)(a)(ii) AND 5(4)(b)(i) AND 5(4)(b)(ii).
Range of elements that the vectors may contain
Vectors may contain regulatory elements, coding or non-coding genes, and regulatory regions of genes.
Regulatory elements:
• Promoters (constitutive, endogenous or inducible) (e.g. chicken β-actin promoter, β-tubulin promoter, rapamycin-responsive promoter, U6 promoter).
• Enhancers (e.g. woodchuck post-transcriptional regulatory element (WPRE)).
• Internal ribosome entry site (e.g. encephalomyocarditis virus IRES).
• Regulatory peptides (e.g. 2A peptide).
• Regulatory elements for inducible expression (e.g. rapamycin system).
• Polyadenylation signals (e.g. bovine growth hormone polyA).
• Multiple cloning sites.
• Origins of replication.
• Splice acceptor/donor sites.
• Transcriptional activators.
• Transcriptional terminator sequences.
• Secretory and targeting signals.
• Recombination sites and flanking sequences (e.g. CRE/Lox system).
• Selection markers (e.g. zeocin resistance, kanamycin).
• Insulators.
• Reporters (e.g. enhanced green fluorescent protein, luciferase, secreted alkaline phosphatase).
Coding, non-coding or regulatory regions of genes expressed during animal development, and/or with biotechnological application.
Including:
• Flanking sequences for targeted homologous recombination.
• Variants with nucleotide substitutions or deletions to determine functional domains or to modify activity.
• Truncations or short inverted sequences (RNAi) as inhibitors.
• Addition of commercially available protein tags (e.g. his tag) to determine transgene localisation and/or expression.
Type, source and function of any donor genetic material
Genetic material may include cDNA or genomic DNA sequences and may be sourced from the Kingdoms Animalia, Planta, Fungi, Protista and Monera and from viruses or viroids.
The donor genetic material will exclude:
• Genetic material, other than that required for lentiviral production, that increases the pathogenicity, virulence, or infectivity of the host organism.
• Genetic material that results in the modified organism having a greater ability to escape from containment than the unmodified host.
• Genes that encode for toxins with an LD50 < 100 µg/kg.
• Genetic material sourced from New Zealand indigenous fauna and flora, Māori, CITES species, or directly sourced human genetic material. Human genetic sequences will be synthesised or obtained from a reputable organisation.
• Nucleic acid sequences coding for a product that can lead to uncontrolled mammalian cellular proliferation.
4.2. Proposed containment of the new organism(s) (physical and operational)
• State which Containment Standard(s) your facility is approved to.
• State the minimum containment level (PC1 or PC2 as per AS/NZS2243.3:2002) required to contain the GMOs (as per the HSNO (Low Risk Genetic Modification) Regulations).
• Discuss whether controls in addition to the requirements listed in the Standard(s) are necessary to adequately contain the GMOs.
VUW operates laboratory containment facilities on the Kelburn campus and at Wellington Hospital which are approved to PC1 and PC2 levels. VUW laboratory facilities are
approved to MAF/ERMA New Zealand Standard Facilities for Microorganisms and Cell Cultures: 2007 for work requiring laboratory containment involving bacteria, cultured cells and embryos.
The VUW small animal facility is approved to MAF/ERMA New Zealand Standard Containment Facilities for Vertebrate Laboratory Animals for work requiring animal containment involving live animals (including recipients carrying genetically modified embryos).
Work carried out in laboratory containment will be undertaken in facilities approved to at least the required physical containment level.
Genetically modified embryos will be transferred to a PC2 vertebrate laboratory animal containment facility (as described in MAFBA Std. 154.03.03 and 2243.3:2002) for implantation into recipient animals (mice and rats only). Embryos will be transferred between containment facilities in a double enclosed container.
5. Risks, costs and benefits
Provide information of the risks, costs and benefits of the GMOs in the following areas of impact:
• The environment.
• Human health and safety.
• The economy (e.g. the ability of people and communities to provide for their economic wellbeing).
• The relationship of Māori and their culture and traditions with their ancestral lands, water, sites, waahi tapu, valued flora and fauna and other taonga, and the principles of the Treaty of Waitangi (The details of any engagement or consultations with Māori that you have undertaken in relation to this application should be discussed here).
• Society and community.
• New Zealand’s international obligations.
What adverse effects could this organism have on the environment?
For all stages of the life cycle
All organisms that will be developed in this project meet the low-risk regulations and are kept in physical containment. Thus, it is expected that these organisms will have minimal environmental risks associated with them. Even in the unlikely event of escape into the environment, environmental risks are minimal as further outlined below.
The attenuated strains of E. coli used to propagate the vector packaging plasmids and for cloning are highly unlikely to survive outside of laboratory conditions, thus any adverse effect on the environment is highly improbable.
The mammalian cells lines transduced with lentiviral particles or transfected with lentiviral packaging and expression expression plasmids require stringent culture conditions and will therefore not survive outside of laboratory conditions, thus any adverse effect on the environment is highly improbable. Mammalian embryos have even stricter media requirements and are highly labile, posing less risk than cell lines..
The potential for lentiviral particles to survive outside of containment and to transduce animals in the environment is extremely low; therefore, inadvertent release of lentiviral particles would have negligible environmental impact. Deliberate innoculation of high titer lentivirus particles to animals outside of containment would have negligible environment impact considering that 1) the lentivrus is replication defective and 2) the packaged genetic material will not result in increased pathogenicity, infectivity, virulence or toxicity nor does it contain genes causing an increase in cell proliferation.
Animals genetically modified with lentiviruses will be kept in containment. In the unlikely event that GMOs escaped, or were released, from MAF-approved containment facilities it is highly unlikely that the GMO would have any more adverse effect on the environment than an unmodified host.
Recipients animals receiving genetically modified (via lentiviruses) embryos are at minimal risk of being transduced with lentivirus because (i) the virus particles remaining in the medium are labile and decay and (ii) embryos will have been washed several times. A high titer is imperative
for transduction. Should individual uterine cells of recipients become transfected the replication- deficient nature of the virus will ensure no spreading so the effect would be negligible.
The ability to produce a replication competent vector by non-homologous recombination is eliminated by the design of the lentiviral system, as previously explained.
The generation of replication competent vector through super infection of lentivirus transduced cells with wild type lentiviruses is unlikely adue to the safety modifications previously listed but is theoretically possible. In the unlikely event that replication competent lentivirus was generated it would have negligible environmental impact since (1) still require escape or deliberate release of animals from containment and (2) as per the exclusions, packaged genetic material will not result in increased pathogenicity, infectivity, virulence or toxicity.
The animals to be used in this application are already common in New Zealand and hence there is no risk of introducing a new species to the environment.
What adverse effects and benefits could this organism have on human health and safety?
The genetically modified organisms would pose no risk to human health or safety.
The greatest risk of occupational exposure to staff would be during the preparation of viral particles. The use of Biological Safety Cabinets during production will minimize this possibility.
Even with deliberate or inadvertent injection of high titer lentiviral vectors the exclusion of genetic elements that would result in increased pathogenicity, infectivity, virulence or toxicity would minimize the possibility of adverse effects on human health.
The genetically modified animal models would have a benefit on human health by contributing to research aimed at improving health and combating disease.
What adverse economic effects and benefits could this organism have?
To the best of our knowledge, there are no economic effects that inadvertent or deliberate release of the genetically modified organisms described in this application would entail.
Benefits include reduction in health costs via research leading to new treatments.
What adverse effects could this organism have on the relationship of Māori and their culture and traditions with their ancestral lands, water, sites, waahi tapu, valued flora and fauna and other taonga (taking into account the principles of the Treaty of Waitangi)?
Include details of any consultation that you have undertaken.
To the best of our knowledge, there are no adverse effects of the GMOs on the relationship of Māori and their culture and traditions, Genetic material will not be sourced from New Zealand indigenous fauna and flora and human genetic material will be obtained from reputable commercial suppliers or research institutes and will not be derived from Māori donors. In addition, the work is undertaken in physical containment minimising any potential environmental impact.
What adverse effects on society, communities and New Zealand’s international obligations could this organism have?
To the best of our knowledge, there are no adverse effects that inadvertent or deliberate release of the genetically modified organisms described in this application would entail.
6. Other information
Add here any further information you wish to include in this application including if there are any ethical considerations that you are aware of in relation to your application.
7. Checklist
This checklist is to be completed by the applicant
Application Comments/justifications
All sections of the application form completed or you have requested an information waiver under section 59 of the HSNO Act
☐
Yes☐
No(If No, please discuss with an Advisor to enable your application to be further processed)
Confidential data as part of a separate, identified appendix
☐
Yes☐
NoSupplementary optional information attached:
• Copies of additional references
☐
Yes☐
No• Relevant correspondence
☐
Yes☐
No AdministrationAre you an approved EPA customer?
☐
Yes☐
NoIf Yes are you an:
Applicant:
☐
Agent:
☐
If you are not an approved customer, payment of fee will be by:
• Direct credit made to the EPA bank account (preferred method of payment) Date of direct credit:
• Cheque for application fee enclosed
☐
Yes☐
No☐
Payment to follow☐
Yes☐
No☐
Payment to follow
Electronic, signed copy of application e- mailed to the EPA
☐
YesSignature of applicant or person authorised to sign on behalf of applicant
☒
I am making this application, or am authorised to sign on behalf of the applicant or applicant organisation.☒
I have completed this application to the best of my ability and, as far as I am aware, the information I have provided in this application form is correct.Peter Pfeffer 8 April 2022
Signature Date
Request for information waiver under section 59 of the HSNO Act
☐
I request for the Authority to waive any legislative information requirements (i.e. concerning the information that has been supplied in my application) that my application does not meet (tick if applicable).Please list below which section(s) of this form are relevant to the information waiver request:
Appendices and referenced material (if any) and glossary (if required)
Glossary of scientific and technical terms used in the application
Expression construct – a DNA molecule containing a combination of coding and regulatory sequences necessary to produce expression of a transgene
Gene targeting – site-specific genetic modification of chromosomal DNA by homologous recombination
Gene transfer - the extra-chromosomal expression of a gene expression construct that doesn’t interfere with the integrity of the host genome
Homologous recombination – genetic recombination between two identical strands of DNA.
In ovo – “within the (chicken) egg” achieved by cutting a sealable window into the egg shell through which the embryo can be seen and accessed
Non-conjugative bacterial strain – strains that are unable to mediate the transfer of genetic material (plasmids) to other bacteria.
Promoter – piece of DNA that drives the expression of a transgene.
Reporter gene – a gene with a product that can be readily detected or that catalyses a reaction that can be readily detected
RNAi – various methods of down regulating RNA expression by small RNA molecules.
Somatic – non-germline tissues.
Transduction – the transfer of the genetic material into a target cell by a recombinant viral vector.
Transfection – transfer of genetic material into a host cell, usually in a plasmid vector.
Transgene – the selected gene-of-interest or genetic material.
Transgenic – genetically modified organism. Organism whose genome has been altered by the inclusion of foreign genetic material.
