New York UniversityRoslin Institute/ University of EdinburghMichigan State University
Transgenic approach to improved productivity:Establishing African Trypanosomiasis resistance in cattle by a
transgenic approach
Mingyan Yu, Charity Muteti, Moses Ogugo, Steve KempAnimal Biosciences, ILRI
ILRI BioSciences Day, Nairobi, 27 November 2013
Partner go
Trypanosomes in Blood stream
African Trypanosomiasis• Caused by extracellular protozoan
parasites – Trypanosoma• Transmitted between mammals by
Tsetse flies (Glossina sp.)• Prevalent in 36 countries of sub-Sahara
Africa.
In cattle• A chronic debilitating and fatal disease.• A major constraint on livestock and
agricultural production in Africa. • Costs US$ 1 billion annually.
In human (Human Sleeping Sickness)• Fatal• 60,000 people die every year• Both wild and domestic animals are
the major reservoir of the parasites for human infection.
http://www.bio.davidson.edu/courses/immunology/students/spring2006/ryan/termpaper.html
http://www.imib-wuerzburg.de/research/siegel/research/
The problem
Vector Control (Tsetse Fly)• Using toxic insecticide• Negative impacts on environment• Not sustainable• Expensive
Vaccine• Tryps periodically change the major surface antigen – variant
surface glycoprotein (VSG) and evade the host immune system.• More than 2 decades, there is no effective vaccine developed.
Drug• No prophylactic drugs• Drug toxicity and resistance• Expensive
New idea
The problem
• Establish a transgenic cattle model with African Trypanosomiasis resistance using nuclear transfer (cloning).
• On the background of a Kenyan indigenous breed – Kenyan Boran.
• Introduce the gene – apoL-I from Baboon into Boran, which is the key trypanolytic component of Baboon’s protective Trypanosome Lytic Factor (TLF) against both cattle and human-infective trypanosomes.
Establish African Trypanosomiasis Resistance in Cattle by A Transgenic Approach
Innovation in the work
• Using a transgenic approach to create disease resistance in cattle
• The method is once for all and self-sustainable - Once the resistance is established, it could be transmitted to the next generations through normal breeding.
6
Links to ILRI’s SO and CGIAR SLO
Tryps Resistant Cow
Improved livestock productivity
Improved crop productivity
More meat and milk
More income from sales
Small-scale
Farmers
SLO1Reduce rural
poverty
SLO3Improve nutrition
and health
SLO2Improve food
security
CGIAR System Level Outcomes
CRP 3.7 – Genetic
component
ILRI SO:Better lives
through livestock
Delivery partners needed
7
Importance to ILRI
• Improved capacity with new technology (cloning) platform
• Being the lead in transgenic livestock research in Africa (challenge & opportunity)
• Improved public visibility and image by resolving the persistent trypanosomiasis problem
Project Process
Genomic locus of Baboon apoL-I gene
Vector construction
Validate the construct in transgenic mouse
Bovine embryonic fibroblasts (BEF) primary culture
Transfection & screening
apoL-I Transgenic BEFs (male)
Nuclear Transfer
(cloning)
Transgenic calves
Phenotyping
Trypanosome resistant transgenic Boran bull
ILRI
ILRI
Kenya Boran
Roslin Institute
New York University
Michigan State University
ID: TatuDate of Birth: 16 July 2012 (Kapiti)Sex: MaleBirth Weight: 46 kgDate of Death: 19 July 2012 (74 hrs)Cause of death: Low temperature, low blood glucose …
ID: CL001 (Tumaini)Date of Birth: 21 August 2012 (ILRI)Sex: MaleBirth Weight: 35 kgCurrent age: 15 months, healthy
ResultsTwo Cloned Calves born through Caesarean Section
At B
irth
15-M
onth
CL001 (Tumaini)
This work is funded by US National Science FoundationBREAD Program
It is implemented in a partnership with New York UniversityMichigan State UniversityRoslin Institute, University of Edinburgh
Fund & Partnership
RDA-ILRI Fund(Korean)&
Transfection of Boran BEFs line (Roslin Institute, UK)
Establish Apol-I Transgenic Boran by nuclear Transfer with Transgenic Cells
Phenotyping (confirm Tryps resistance)
• ApoL-I expression pattern
• Killing of Trypanosomes in vitro
(serum) and in vivo (challenge)
• Monitor the health conditions with
growth
Increase Genetic Diversity• Establish more transgenic cattle
with Kenya Boran BEFs lines• Establish transgenic cattle with
other Kenyan indigenous breeds
Transgene Delivery• Develop a breeding programme to
disseminate the transgene to farmers
Current Project Potential Future Projects
Biosafety Research• Confirm no health risk of the
gene to the transgenic cow• Confirm the safety of meat and
milk consumption of the apoL1 transgenic cow
Where to from now?
Potential opportunities
More application of established cloning platform
• Improve cattle genetics through traits introduction between breeds
- Using TALENs and cloning together to introduce good traits between breeds to obtain efficient cross-breeding.
• Indigenous breeds conservation - Establish a bank of cells from indigenous breeds of cattle, sheep and goats, which are capable of cloning.
• Training for other African regions - Make good use of the technique to maintain the best animal genetics and improve the livestock productivity in Africa.
Resource mobilization strategy• Trypanoresistant cow project – wait and see• Traits introduction between breeds using TALENs -
seeking collaboration and new traits
What support do you expect from ILRI and the BioScience Directorate?
• Effective communications with the public regarding the transgenic cow project in due course.
General discussion (facultative)
The presentation has a Creative Commons licence. You are free to re-use or distribute this work, provided credit is given to ILRI.
better lives through livestock
ilri.org