Transgenic approach to improved productivity: Establishing African Trypanosomiasis resistance in cattle by a transgenic approach

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Published on January 28, 2014

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Presented by Mingyan Yu, Charity Muteti, Moses Ogugo and Steve Kemp at the ILRI BioSciences Day, Nairobi, 27 November 2013

Transgenic approach to improved productivity: Establishing African Trypanosomiasis resistance in cattle by a transgenic approach Mingyan Yu, Charity Muteti, Moses Ogugo, Steve Kemp Animal Biosciences, ILRI ILRI BioSciences Day, Nairobi, 27 November 2013 Partner go New York University Roslin Institute/ University of Edinburgh Michigan State University

The problem African Trypanosomiasis • Caused by extracellular protozoan parasites – Trypanosoma • Transmitted between mammals by Tsetse flies (Glossina sp.) • Prevalent in 36 countries of subSahara Africa. http://www.imib-wuerzburg.de/research/siegel/research/ Trypanosomes in Blood stream 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

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

Establish African Trypanosomiasis Resistance in Cattle by A Transgenic Approach • 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 humaninfective trypanosomes.

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.

Links to ILRI’s SO and CGIAR SLO Tryps Resistant Cow Delivery partners needed ILRI SO: Better lives through livestock Small-scale Farmers Improved livestock productivity More meat and milk More income from sales SLO3 Improve nutrition and health Improved crop productivity SLO2 Improve food security CRP 3.7 – Genetic component SLO1 Reduce rural poverty CGIAR System Level Outcomes 6

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 7

Project Process Kenya Boran Genomic locus of Baboon apoL-I gene Vector construction ILRI Bovine embryonic fibroblasts (BEF) primary culture Roslin Institute Validate the construct in transgenic mouse Transfection & screening apoL-I Transgenic BEFs (male) Nuclear Transfer ILRI (cloning) Transgenic calves Phenotyping Trypanosome resistant transgenic Boran bull New York University Michigan State University

Results Two Cloned Calves born through Caesarean Section ID: Tatu Date of Birth: 16 July 2012 (Kapiti) Sex: Male Birth Weight: 46 kg Date 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: Male Birth Weight: 35 kg Current age: 15 months, healthy

15-Month At Birth CL001 (Tumaini)

Fund & Partnership This work is funded by US National Science Foundation & BREAD Program RDA-ILRI Fund (Korean) It is implemented in a partnership with New York University Michigan State University Roslin Institute, University of Edinburgh

Where to from now? Current Project Potential Future Projects Transfection of Boran BEFs line (Roslin Institute, UK) 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 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

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.

General discussion (facultative) 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.

better lives through livestock ilri.org The presentation has a Creative Commons licence. You are free to re-use or distribute this work, provided credit is given to ILRI.

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