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