Regulations of the Genome Engineering Unit (hereinafter referred to as “GEU”)
- GEU has been established with the support from the Foundation for Polish Science under the project called “Mouse Genome Engineering Facility – Generation of Animal Models for Biomedical Research and Preclinical Studies TEAM-TECH CORE FACILITY”, as part of the Innovative Development Operational Programme (POIR.04.04.00-00-436A/17-00).
- GEU is an organizational unit within the IIMCB, supervised by the Head of the Unit, overseen by the IIMCB Director
- Scientific advisor for GEU is Prof. Andrzej Dziembowski, Group Leader of the Laboratory of RNA Biology – ERA Chairs Group.
- GEU has been established to render scientific services for employees and doctoral students of IIMCB, other entities whose main objective is to conduct scientific research as well as commercial entities. The provided services are:
a) generation of transgenic mouse models for biomedical research and preclinical studies,
b) other embryological services (e.g. cryopreservation of embryos, in vitro fertilisation),
c) preparation of plasmids,
e) other molecular biology services.
- GEU renders its services by employing top professionals in molecular biology and developmental biology and has an access to highly-specialized scientific and research equipment operated only by a team of adequately trained employees and doctoral students.
- Within the scope of animal studies, GEU operates based on valid permissions from the Local Ethical Board (hereinafter referred to as “LEB”) while the staff working with bred animals (mice) have been individually licensed to perform specific procedures on animals.
- No person without appropriate training.is allowed to use the equipment being at the disposal of GEU
- IIMCB, on behalf of GEU, provides the services under the relevant agreements:
a) on access to the breeding area in a specialized animal lab that meets all the requirements defined in the Act on Protection of Animals Used for Scientific or Educational Purposes dated 17 January 2021 (Journal of Laws of 2021, item 2338) and the Regulation of the Minister of Agriculture and Rural Development on minimum requirements that should be met by laboratories and minimum requirements of lab animal care dated 14 December 2016 (Journal of Laws of 2016, item 2139);
b) license agreements on the use of patented CRISPR/Cas9 technology.
- The settlement of transactions connected with the activity of GEU is controlled and approved by the IIMCB Director in compliance with the principles followed at IIMCB.
General rules for services
- GEU renders the services specified in § 1 sec. 4 and § 3 sec. 1 of these Regulations.
- The services are rendered in compliance with applicable regulations, instructions from funding institutions and, in scope for mouse line generation, provisions outlined in the Agreement on Execution of the TEAM TECH CORE FACILITY Project(POIR.04.04.00-00-436A/17-00).
- For transgenic mouse line generation, GEU ensures equal access to the services for all potential recipients. The difference in price between academic and commercial users (hereinafter referred to as Ordering Parties) is solely due to the different levels of fees independent of IIMCB.
- Price list for other services is prepared according to the rules stated in Order no 4/2022 dated 8 April 2022
- The performance of service commences on the date of acceptance of an order confirmed via e-mail by the Head of GEU (in case of mouse model generation) or an employee of GEU (other orders). The duration of service performance is determined on a case-by-case basis
- Settlements are made based on VAT invoices issued to external entities after the receipt of the service or an accounting order in the case of services rendered for internal recipients, employees carrying out projects at IIMCB
- The cost of transportation of the mice ordered, additional insurance costs, additional fees (e.g. custom duties) shall be borne by the Ordering Party.
Detailed rules for mouse line generation service
1. GEU offers a wide variety of genetic modifications in the mouse genome, including among others:
a) introduction of indel mutations in the desired locus in mice,
b) introduction of site-specific deletion or point mutation in the desired locus in mice,
c) construction of knock-in mice,
d) construction of mouse lines expressing Cre recombinase in specific tissues,
e) introduction of site-specific point mutation: disease-related mutations, catalytic mutations, etc.,
f) generation of specific conditionally regulated allele, including conditional knock-out, in which LoxP site is introduced to introns surrounding a relevant exon.
2. Orders will be executed after appropriate approvals have been obtained from LEB. LEB applications will be submitted by the IIMCB. The Ordering Party is obliged to prepare a project description containing information necessary for the preparation of the application to the LEB
3. Predicted mouse phenotypes have to be discussed with the Head of GEU before the start of the project, especially potential harmful phenotype or phenotypes leading to embryolethality or infertility have to be disclosed.
4. After submitting the order, the Ordering Party will receive the mutagenesis strategy for acceptance. Once accepted, no change to the strategy may be made without the written consent of both parties. GEU does not take responsibility for the physiological effect of the introduced mutations.
5. The price list for mouse line generation is approved by the Director of IIMCB and published on the GEU website (crisprimice.eu), IIMCB (www.iimcb.gov.pl) and on the IIMCB intranet
6. For all projects, GEU will make at least 3 attempts for mouse line generation.
7. All mouse lines will be generated till N1 generation (after 1 crossing of founder mouse with WT mouse). Ordering Party will only be charged for a successful mouse line generation, with the following exceptions:
a) if no potential harmful phenotype was disclosed and F0 mutants die or display a severe phenotype leading to the implementation of humane endpoint – Ordering Party will be charged full price. The experiment can be repeated only if the Ordering Party receives appropriate approval by the LEB,
b) where the possibility of a harmful phenotype has been reported, individual arrangements between the Ordering Party and the Head of the GEU will apply,
c) If heterozygous F0 mice of both sexes are infertile, in vitro fertilization will be attempted. If in vitro fertilization proves unsuccessful, F0 generation mice will be transferred to the Ordering Party. The Ordering Party will be charged the full amount,
d) In case of generation of knock-out or point mutation mouse lines: if heterozygotic mice prove embriolethal and potential embriolethality was not disclosed, Ordering Party will be charged 60% of the price. Mouse line can be considered embryolethal if planned mutation is present in >20% of blastocysts and no mice with the correct mutation are born after the successful transfer of at least 100 blastocysts. Where the possibility of embriolethality of heterozygotic mutants has been reported, individual arrangements between the Ordering Party and the Head of the GEU will apply.
8. Upon completion of the project, the customer shall receive:
a) 5-6 weeks old N1 generation mice (at least 1 breeding pair, in most cases 5-10 mice),
b) map of the mutation introduced (Snap Gene file),
c) sequencing chromatograms of N1 mice,
d) genotyping protocol,
e) health report according to Felasa guidelines (current and historical data for 24 months),
f) data on observed phenotype related to the mutation (if any), g) upon request: health report signed by the veterinarian
9. Considering the welfare of animals, GEU reserves the right to suspend deliveries of animals in extreme weather conditions. Should this be the case, another delivery date will be agreed upon with the Ordering Party.
10. The Ordering Party commits to cryopreserve the received mouse line in the form of embryos or sperm. Cryopreservation may be outsourced to GEU for an additional fee.
11. Considering the welfare of animals, GEU reserves the right to suspend deliveries of animals in extreme weather conditions. Should this be the case, another delivery date will be agreed upon with the Ordering Party.
Detailed rules for cloning service
1. GEU offers preparation of plasmids according to Ordering Parties' needs, based on a broad range of backbone vectors.
2. To order a plasmid, the Ordering Party should contact a GEU employee providing the following information:
a) Insert sequence,
b) Source of the template to amplify the insert,
c) Backbone vector,
d) C- or N-terminal tag
3. The estimated time for vector preparation is 1 month.
4. Ordering Parties will only be charged for successful clonings, with the following exceptions:
a) if wrong insert sequence or template for insert amplification is provided, Ordering Party is charged the full price, b) if cDNA is used as a PCR template and no PCR product is produced after 4 attempts in various conditions – project will be aborted and the Ordering Party will be charged a fee specified in the price list,
c) in case of cloning from Danio rerio cDNA: cloned sequence usually does not 100% match the reference sequence (several point mutations are present). This will not be considered unsuccessful cloning. Mutations can be corrected to match the reference sequence for an additional fee specified in the price list.
5. After project completion Ordering Party will be provided:
a) at least 0,5 ug of midi-prep purified plasmid,
b) map of the plasmid (Snap Gene file),
c) sequencing chromatograms.
6. GEU will keep plasmid for at least 1 year
Should the results achieved on the basis of the services rendered at GEU be published, information on the location of service performance should be provided (e.g. “Research model/Plasmid was generated by the Genome Engineering Unit of the International Institute of Molecular and Cell Biology in Warsaw”).
1. The Head of the GEU shall be responsible for training the staff of GEU as regards equipment handling and security.
2. The Head of the GEU shall be responsible for training the staff and/or shall have training provided to the staff performing procedures on animals.
2. Should a service have a physical defect, GEU shall render such service once more or – at its discretion – refund the cost of the service.
3. GEU waives all implied guarantees, including implied guarantees of marketability or fitness for a specific purpose, or guarantees of non-violation of intellectual property rights.
4. GEU shall only be liable if any damage results from the Facility’s wilful act or gross negligence of staff performing the service and only up to the value of the actual loss, subject to sec. 5 below.
5. The liability of GEU shall be limited to the price of the service.
1. Upon placement of an order, the Ordering Party accepts the Regulations of GEU.
2. The Parties undertake to solve any disputes resulting from or related to these Regulations amicably, in mutual negotiations.
3. If the Parties fail to reach an agreement within 14 days from the onset of a dispute, it shall be solved by the competent court of law for the registered office of GEU.