chIFITM knockdown/knockout technology as a platform technology for increased vaccine yields.
Lead Research Organisation:
The Pirbright Institute
Department Name: Pirbright Laboratory
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
People |
ORCID iD |
| Mark Fife (Principal Investigator) |
Publications
Bassano I
(2017)
Accurate characterization of the IFITM locus using MiSeq and PacBio sequencing shows genetic variation in Galliformes.
in BMC genomics
| Description | This Funding provided essential data for a successful application to the Bill and Melinda Gates foundation. |
| Exploitation Route | Numerous vaccines are produced in hen's eggs or cell lines, but the amount that can be produced is limited by the immune responses that prevent replication of vaccine viruses. The funding will enable previous research carried out by the Genetics and Genomics group at Pirbright to be continued. The researchers were the first to describe a set of immune proteins in the chicken called chIFITMs, which prevent viruses from multiplying in cells. Many vaccines, for both animal and human, are produced by growing a weakened form of the virus in chicken eggs or cells, which are then extracted for use. Although chIFITM may help protect chickens from viral infection, the protein actually hinders vaccine production, as it prevents the weakened virus from replicating at high levels and reduces the amount of vaccine that can be made. Our new research will involve using a gene editing system called CRISPR/Cas9 to remove the chIFITM genes in chicken cells, therefore overcoming one of the barriers for viral replication, and boosting the levels of vaccine virus produced. For example, the flu vaccine currently requires two eggs to produce a single dose, but inactivating chIFITM genes could mean only a single egg is needed per dose. This increase in yield will make vaccines cheaper to produce and more accessible to livestock owners in developing nations, the main target of this funding. Initially the scientists will focus on increasing flu vaccine yields, but the technique can be applied to multiple livestock viral diseases and potentially human diseases too. Once the researchers have piloted this technique, for which a patent has been filed, they will work with commercial partners to bring this new technology to market. By 2024, global poultry meat consumption is expected to rise by over 20 million tonnes compared to 2015, and it is therefore essential to maximise outputs worldwide. Poultry production is persistently reduced through viral infection; developing efficient and affordable vaccines against viral diseases will alleviate poverty in developing countries, where these diseases cause devastating consequences for subsistence poultry farming. This research has the potential to increase vaccine yields between five and ten-fold, which, in an industry where the total revenue for vaccines produced in eggs and cell lines is approximately $14.3 billion, can make vast differences to manufacturing costs which will in turn enable vaccine prices to be reduced. The project is set to run for four years, so this new technology could be commercially available as early as 2021. |
| Sectors | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology |
| URL | https://www.pirbright.ac.uk/our-science/avian-viral-diseases/genetics-and-genomics |
| Description | Numerous vaccines are produced in hen's eggs or cell lines, but the amount that can be produced is limited by the immune responses that prevent replication of vaccine viruses. The funding will enable previous research carried out by the Genetics and Genomics group at Pirbright to be continued. The researchers were the first to describe a set of immune proteins in the chicken called chIFITMs, which prevent viruses from multiplying in cells. Many vaccines, for both animal and human, are produced by growing a weakened form of the virus in chicken eggs or cells, which are then extracted for use. Although chIFITM may help protect chickens from viral infection, the protein actually hinders vaccine production, as it prevents the weakened virus from replicating at high levels and reduces the amount of vaccine that can be made. Our new research will involve using a gene editing system called CRISPR/Cas9 to remove the chIFITM genes in chicken cells, therefore overcoming one of the barriers for viral replication, and boosting the levels of vaccine virus produced. For example, the flu vaccine currently requires two eggs to produce a single dose, but inactivating chIFITM genes could mean only a single egg is needed per dose. This increase in yield will make vaccines cheaper to produce and more accessible to livestock owners in developing nations, the main target of this funding. Initially the scientists will focus on increasing flu vaccine yields, but the technique can be applied to multiple livestock viral diseases and potentially human diseases too. Once the researchers have piloted this technique, for which a patent has been filed, they will work with commercial partners to bring this new technology to market. |
| First Year Of Impact | 2017 |
| Sector | Agriculture, Food and Drink,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Societal |
| Description | LVIF |
| Amount | $1,500,000 (CAD) |
| Organisation | Bill and Melinda Gates Foundation |
| Sector | Charity/Non Profit |
| Country | United States |
| Start | 04/2018 |
| End | 09/2020 |
| Description | Pathfinder BB/R012431/1 Grant holder: Dr Mark Fife. Grant title: chIFITM knockdown/knockout technology as a platform technology for increased vaccine yields in SPF Eggs. |
| Amount | £12,500 (GBP) |
| Funding ID | BB/R012431/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 11/2017 |
| End | 02/2018 |
| Title | Comprehensive analysis of genetic variants across immune loci in indigenous breeds. |
| Description | The interferon-induced transmembrane (IFITM) protein family comprises a class of restriction factors widely characterised in humans for their potent antiviral activity. Their biological activity is well documented in several animal species, but their genetic variation and biological mechanism is less well understood, particularly in avian species. Here we report the complete sequence of the domestic chicken Gallus gallus IFITM locus from a wide variety of chicken breeds to examine the detailed pattern of genetic variation of the locus on chromosome 5, including the flanking genes ATHL1 and B4GALNT4. We have generated chIFITM sequences from commercial breeds (supermarket-derived chicken breasts), ancient bone samples, indigenous chickens from Nigeria (Nsukka) and Ethiopia, European breeds and inbred chicken lines from The Pirbright Institute, totalling of 211 chickens. Through mapping of genetic variants to the latest chIFITM consensus sequence our data reveal that the chIFITM locus does not show structural variation in the locus across the populations analysed, despite spanning diverse breeds from different geographic locations. However, single nucleotide variants (SNVs) in functionally important regions of the proteins within certain groups of chickens were detected, in particular the European breeds and indigenous birds from Ethiopia and Nigeria. In addition, we also found that two out of four SNVs located in the chIFITM1 (Ser36 and Arg77) and chIFITM3 (Val103) proteins were simultaneously under positive selection. Together these data suggest that IFITM genetic variation may contribute to the capacities of different chicken populations to resist viral infection. |
| Type Of Material | Model of mechanisms or symptoms - non-mammalian in vivo |
| Year Produced | 2019 |
| Provided To Others? | Yes |
| Impact | This paper is currently in press in. BMC Genomics. |
| Description | Horizon Discovery Ltd Cambridge Research Park, United Kingdom |
| Organisation | Horizon Discovery Group plc |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Verify IFITM knock-out cell by qPCR and immuno-blotting. Verification of further knock-out cell lines by DNA sequencing, immuno-blotting and qPCR Cell infection with Avian Influenza Viruses Assess the viral titre of cells infected with Influenza Viruses in K/O and wildtype cells. Quantitative analysis of viral infection in IFITM K/O cells Demonstrate increased AIV viral titres at lab scale and estimation of what this may equate to with respect to vaccine dose yields. Analysis of IFITM K/O cell lines permissivity to a range of additional animal viruses (Relevant to the LVIF). Milestone 7: Demonstrate permissivity increased and viral titres for a range of LVIF relevant viruses in the edited cell lines. |
| Collaborator Contribution | Develop a CRISPR/cas9 gene editing system to specifically target the chIFITM locus in avian cell culture |
| Impact | Grant funding |
| Start Year | 2017 |
| Description | ILRI Research Collaboration (Kenya, March 2017) |
| Organisation | International Livestock Research Institute (ILRI) |
| Country | Kenya |
| Sector | Charity/Non Profit |
| PI Contribution | Exchange of ideas and samples for indigenous breeds of livestock |
| Collaborator Contribution | Contributed samples for analysis. |
| Impact | Analysis of indigenous breed genomic sequences for Immune function genes. |
| Start Year | 2017 |
| Description | Industrial partnership-Aviagen 2018 |
| Organisation | Aviagen Group |
| Country | United States |
| Sector | Private |
| PI Contribution | We have established a significant industrial partnership with Aviagen group Ltd. The nature of this work remains confidential. |
| Collaborator Contribution | The nature of this work remains confidential. |
| Impact | The nature of this work remains confidential. |
| Start Year | 2018 |
| Description | Research Collaboration with Nigeria (23rd - 28th January 2017) |
| Organisation | University of Nigeria |
| Country | Nigeria |
| Sector | Academic/University |
| PI Contribution | Analysis of rare breed indigenous chicken in Nigeria. |
| Collaborator Contribution | Helped with sampling and background information of the breed. |
| Impact | Analysis is ongoing. |
| Start Year | 2017 |
| Title | AVIAN CELLS FOR IMPROVED VIRUS PRODUCTION |
| Description | The present Invention provides as avian cell in which the expression or activity of one or more of the following genes, or a homologue thereof: Chicken IFITM 1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3) is reduced. The invention also provides methods for passaging viruses in avian cells, embryos and/or avian cell lines which have reduced expression of one or more IFITM genes and methods which involve investigating the sequence of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3). |
| IP Reference | WO2014195692 |
| Protection | Patent granted |
| Year Protection Granted | 2014 |
| Licensed | Commercial In Confidence |
| Impact | The present invention provides an avian cell in which the expression or activity of one or more of the following genes, or a homologue thereof: Chicken IFITM1 (SEQ ID No. 1); Chicken IFITM2 (SEQ ID No. 2) and Chicken IFITM3 (SEQ ID No. 3) is reduced. The invention also provides methods for passaging viruses in avian cells, embryos and/or avian cell lines which have reduced expression of one or more IFITM genes and methods which involve investigating the sequence of one or more of the follow |
| Description | Chair of organising Committee for Wellcome Trust-Animal Genetics and Diseases 2017 |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | This meeting brought together specialists working on the interface between genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare. Scientific sessions included: Genetics of immune responses and disease resistance Genetically engineered livestock (including genome editing) Quantitative genetics and epigenetics applied to disease Epidemiology and pathogen evolution Bioinformatics, comparative and functional genomics Precision medicine of animal companions |
| Year(s) Of Engagement Activity | 2017 |
| URL | https://coursesandconferences.wellcomegenomecampus.org/events/item.aspx?e=635&dm_i=2SUU,HOGH,4R4AW1,... |
| Description | Encouraging women into science and engineering STEM. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | MH Visited Warwick School, Redhill to talk to secondary students about careers in STEM. A very positive outcome and well received. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Holt School Surrey. Bee meadow & A-level outreach Talk. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | I discussed the importance of pollinators for biodiversity and food security with the younger students. I also discussed careers in science with A-Level students. There was good interaction and many questions surrounding both topics. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Kingdown School Warminster Wiltshire talk to 6th form students about career in science. |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | I presented my career path to the students to engage them on STEM careers. I had a good level of discussion and many questions. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Organiser of the Animal Genetics and Diseases conference 08 - 10 May 2019 Wellcome Genome Campus, UK. Highlighting recent advances in animal genetics and genomic technologies. |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | I am on the organising committee for this conference. The second conference in this series will highlight recent advances in animal genetics and genomic technologies. It will bring together specialists working on the interface of genomics, genetic engineering and infectious disease with the aims of improving animal and human health and welfare. Novel genomic technologies, mathematical modelling and quantitative genetics approaches, applied to host animals, as well as their pathogens, have transformed the understanding of animal diseases, host-pathogen interactions and epidemiology and their effects on productivity of farmed animal species and food supply chains. This year's conference will not only put the spotlight on the immune response of host animals and epidemiology but also cover the genetics and genomics of pathogens and the impact of animal-human relationships. We encourage registrations from researchers, breeders and technical specialists interested in learning and disseminating the latest cutting-edge techniques and methodologies across model species, wildlife, farmed animals and companion animals. |
| Year(s) Of Engagement Activity | 2019 |
| URL | https://coursesandconferences.wellcomegenomecampus.org/our-events/animal-genetics-diseases-2019/?dm_... |
| Description | Taiwan -UK Partnering Award: Surveillance of influenza viruses |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Attended the International Flu Virus Symposium for Celebrating IPM 47th Anniversary" in Tri-Service General Hospital in Taipei, Taiwan. Tri-Service General Hospital address: No. 325, Sec. 2, Chenggong Rd., Neihu Dist., Taipei City 114, Taiwan (R.O.C.) Invited speaker for medical staff and research scientists. |
| Year(s) Of Engagement Activity | 2019 |
| URL | https://www.cdc.gov.tw/rwd/english |
| Description | Winston Churchill Careers Fair. Winston Churchill School, Hermitage Road, St Johns, Woking, Surrey, GU21 8TL |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | AS attended and presented at this event to stimulate increased interest in science and research. |
| Year(s) Of Engagement Activity | 2018 |