Dengue,

Mosquitoes
& Genes
An information pack to
accompany the Oxitec film

CONTENTS

1.0

Introduction

2.0

Dengue Fever a growing problem

2.1

Who is at risk?

2.2

Why are they at risk?

2.3

How can we treat it?

3.0

Introducing Haedes and Aegypta: all about the Aedes aegypti mosquito

3.1

Dining out with Aegypta: how and why do mosquitoes suck blood?

3.2

How does the Aegypti mosquito transmit Dengue fever?

3.3

Getting on top of the problem: how we can make life uncomfortable for Aegypta and friends

4.0

Using genes to control insects: the Oxitec solution

4.1

What exactly is a gene, and what is meant by genetics?

4.2

Applying genetic modification to insect control: The Oxitec solution

4.3

Using sterile insects for population control

4.4

The Oxitec approach: genetic engineering of sterile insects

4.5

How can we breed genetically sterile mosquitoes?

4.6

Why genetic modification works

4.7

More on the science: How does Oxitec make genetically modified insects?

Introduction
We hope you enjoy this short film about Dengue
Fever and the Aedes aegypti mosquito.
At Oxitec, were concerned about Dengue Fever because

the disease, and about how Oxitecs cutting-edge

of the devastating impact it has on so many peoples

solution is providing a new approach to controlling them.

lives, and because of the huge costs which Governments

and communities must meet in order to try to control this

Weve designed this information pack to be a

dangerous disease. Thats why were using advanced

supplement to the video, and provide a helpful resource

science, and a process called genetic modification, to

for anyone wanting to learn more about Dengue Fever,

develop a new solution to the Dengue threat by

mosquitoes, and the Oxitec approach.

targeting the mosquitoes which spread it.

As you watch the video, we hope youll learn more about
the Dengue Virus, about the mosquitoes which spread

Dengue Fever a growing problem

Dengue Fever is a dangerous and debilitating disease,

In the Film, youll hear from Hannah Strange and

and its a growing threat to global health. Like Malaria,

Professor Paul Reiter about the ordeal that they suffered

Dengue is spread by a bite from an infected mosquito

when they contracted Dengue Fever. The experiences

(we call diseases of this type mosquito-borne).

they describe are not uncommon.

Although it is not usually fatal, Dengue Fever is an

Unfortunately, for some patients, Dengue Fever can be

extremely serious disease. Dengue symptoms range from

even more dangerous. In the more severe form, known as

mild and flu-like to high fever, rash, severe headache,

Dengue Hemorrhagic Fever (DHF), blood vessels start to

pain behind the eyes, muscle and joint pain. The joint

leak and the blood fails to clot, causing bleeding from

pain can be so severe that Dengue has been given the

the nose, mouth, and gums. Without prompt treatment,

name breakbone fever. Nausea, vomiting, and loss of

the blood vessels can collapse, causing a critical

appetite are also common. These symptoms can last for

condition called Dengue Shock Syndrome. Ultimately,

weeks!

this can lead to fatality: about 25,000 people die from

Dengue Fever every year.

Hannah Strange
Dengue Fever Sufferer

As well as pain and suffering for those people unfortunate enough to catch Dengue, the disease is also a serious financial
burden for the Governments and communities which are struggling to cope with it. Governments spend a lot of money on
efforts to control the spread of the Dengue mosquito, but these efforts are not very effective, as well hear about later. At
the same time, the cost of looking after and treating people affected by Dengue can be huge both for Governments and
the individuals concerned. Here are a few facts about the costs of dealing with Dengue:

2,500,000,000

$1,394

2.5bn people are now estimated to be at risk from

In the Americas and Asia, a study found that the

Dengue

average cost of each hospitalised case was $1,394

25,000

TB+

Between 50-100 million people are infected each

The economic costs of Dengue can be of the same

year, with around 25,000 deaths

order of magnitude as those associated with

Tubercolosis, sexually transmitted diseases

$2,100,000,000
In the Americas, the average economic cost of
Dengue fever is estimated at $2.1bn

(excluding HIV/AIDS), Chagas Disease and

Leishmiasis.

Who is at risk?

Why are they at risk?

After Malaria, Dengue Fever is the second most

The rapid spread of Dengue Fever is due to the global

widespread mosquito-borne disease in the world. The

migration of the mosquitoes which spread the disease.

World Health Organisations has estimated that between

The main culprit is Aedes aegypti, and youll hear more

50 and 100 million people suffer from Dengue Fever

from these in the video! The mosquitoes eggs are

each year: thats more than the population of the UK

extremely hardy: they can survive for months without

every year!

water, allowing them to be transported all over the

world, for example in used tyres and plant containers,

Dengue is also the fastest-growing mosquito-borne

before hatching and infesting a new area.

disease. Since the 1970s, the number of countries

experiencing Dengue outbreaks has grown from 9 to

In the video, youll hear from the scientist Professor Paul

more than 100.

Reiter who explains how the Aedes aegypti mosquito has

spread so rapidly around the world. The life-cycle of a

Today, up to 40% of the worlds population, or 2.5 billion

mosquito is about 3 weeks from hatching, to adult, to

people, is thought to be at risk . Dengue fever occurs in

reproduction. If each female can lay up to 500 eggs, as

most tropical areas of the world. It is common in Asia,

Professor Reiter explains, it is easy to see how quickly a

the Pacific, Australia, Latin America and the Caribbean

new area can become infested!

[1]

and is continuing to spread. It has now reached North

America. A recent Natural Defence Resource Council
report shows that 28 US states are now at risk[2].

[1] World Health Organisation Dengue Factsheet: http://www.who.int/mediacentre/factsheets/fs117/en/index.html

[2] Natural Resources Defence Council (2009) Fever Pitch: Mosquito-borne Dengue Fever Threat Spreading in the Americas http://www.nrdc.org/health/dengue/files/dengue.pdf

How can we treat it?

There is currently no treatment for Dengue fever. One of

Because of the risk of these complications, a new vaccine

the difficulties in developing an effective drug or vaccine

has to be effective against all four types of Dengue virus.

to combat the virus is that Dengue Fever is actually

A vaccine of this type is in development, but it could be

caused by four different, but closely related, types of

many years before it can enter production. Even then, a

virus.

vaccine of this type could still risk actually increasing

peoples sensitivity to one or more Dengue types. Its

Scientists call these related viruses serotypes: the four

important to remember as well that there are an

Dengue serotypes are named DENV1, DENV2, DENV3

estimated 2.5 billion people currently thought to be at

and DENV4. When a person is infected by one of these

risk from Dengue. Vaccines are a safe and effective way

serotypes, they develop life-long immunity to that type,

to control disease, but they are expensive. With so many

but not to the other three. In fact, a person who has been

people at risk from Dengue, some countries may struggle

exposed to one type may be at risk of developing a more

to fund a vaccination programme. If a vaccine

serious illness if they are infected by another type later

programme was implemented, it could be combined

in life. Scientists dont completely understand the reason

with mosquito control strategies to provide even better

for this, but it is likely to be a result of the way our

protection against Dengue.

immune systems respond to the different viruses.

With no drug, and a potential vaccine a long way off, our

only way of controlling Dengue Fever is to target the
mosquitoes which carry it.

Bednets are often used in hospitals within tropical regions, but they are of
limited use in preventing Dengue transmission because the Aedes aegypti
mosquito bites during the day

Introducing Haedes &

Aegypta: all about the
Aedes aegypti mosquito
So youve met Haedes and Aegypta the animated stars
of our film! As you may have noticed, Haedes and
Aegypta get their names from Aedes aegypti, which is
the Latin name for the mosquito which carries Dengue
Fever.
Aedes aegypti is a prolific pest. Originating in Africa, it
has spread around the world, hitching rides in shipping
containers, used tyres, and other transported goods
which can provide ideal vehicles for the mosquitoes
eggs. As Aegypta proudly points out, Aedes aegypti are
highly adaptive: once transported to a new habitat, they
breed quickly and rapidly become established in an area.
As the expert earlier in the video tells us, female Dengue
mosquitoes can produce up to 500 eggs, so Aegyptas
300 children arent unusual!
The Dengue mosquito is primarily an urban pest. That means that it prefers to live in and around human habitation.
It may seem a bit far-fetched to find Haedes and Aegypta setting up home in a fruit bowl, but in fact, thats exactly the
kind of environment which Aedes aegypti mosquitoes prefer. For example, scientists studying the preferred breeding sites
of Aedes aegypti mosquitoes in Singapore City listed a number of common household containers which were frequently
found to contain Aedes aegytpti larvae, or young.
They included:

If you have a quick think about how



often you might find some of these


 



containers lying around in your




own home, you can start to see




how difficult it might be to try to



get rid of Aedes aegypti once it has



become established in an area.







Dining out with Aegypta:

how and why do mosquitoes suck blood?

If youve ever been bitten by a mosquito, youll probably

The images below show how the female mosquito has

be aware that they feed on blood! What you may not

become adapted to take blood meals from people and

have been aware of is that it is only female mosquitoes

animals. In these highly magnified images, it is easy to

which actually do this males dont bite. In the video,

see the females long feeding mouth parts, or proboscis.

youll hear from Haedes and Aegypta about their

In comparison, the males proboscis is much smaller, and

different dietary habits: as Hades explains, hes a

doesnt have some of the specialised mouthparts found

vegetarian: what he means by this is that, like all male

in females. When a female mosquito bites a human or

mosquitoes, he gets his meals from feeding on nectar,

animal, she will insert this proboscis into a blood vessel,

fruit and other sugar sources.

using a back-and-forth motion of her head to create a

sawing motion that drives the piercing parts of the

Female mosquitoes like Aegypta also feed on nectar and

proboscis into the skin. Special chemicals in her saliva

other sugar sources as their primary source of energy.

prevent the blood from clotting, so she can easily suck

However, females need to feed on blood in order for

the blood into her stomach. It is the bodys reaction to

them to produce eggs.

this saliva which causes the area around the mosquito

bite to swell up and itch as most of us will have

What makes Aegypta and the rest of her species

experienced! When a female feeds, her stomach can

especially troublesome to man, and makes her such a

expand to many times its original volume, allowing her

good carrier of dengue and other diseases, is that she

to consume more than her body weight in blood (she can

prefers human blood. Scientists say she is

struggle to fly afterwards!).

anthroprophilic, which means human loving.

Female mosquitoes
mouth parts using a
scanning electron
microscope
A = Labium tip (the labellum); this
guides the mouthparts into the
skin.
B = Labrum; this is the tube through
which the blood is sucked up.
C = Maxilla; these sharp serrated
edges aid penetration of the skin.
D = Hypopharynx; saliva is
delivered through this tube.

How does the Aedes aegypti mosquito transmit

Dengue fever?
Most of us would agree that mosquitoes blood feeding habits can be extremely annoying! But in the case of Aedes
aegypti, its their ability to transmit Dengue Fever that changes them from a mere annoyance into a potentially deadly
threat.
In the video, youll have heard the reporter asking Aegypta and Haedes about Dengue Fever. As Aegypta explains, the
Dengue virus is picked up by the mosquito when they blood feed on an infected person. After infecting the mosquito, the
virus takes about 5-7 days to replicate itself and pass through the mosquitos body eventually reaching its salivary
glands. Once there, it can enter the mosquitos saliva, so when the mosquito bites another person, the virus can pass into
their bloodstream.

Diagram showing the

life-cycle of
Dengue Virus
Once the mosquito has become
infected with Dengue, it remains
that way for the rest of its life. So
every time she bites a human, she
can pass on the deadly virus.

Getting on top of the problem: how we can make

life uncomfortable for Aegypta and friends
Because of the threat posed by the Dengue mosquito,
people have been trying to reduce or eradicate
populations of the pest for many years. Unfortunately,
despite considerable time and money expended by
governments and communities, these efforts have until
now not been very successful.
In the video, Haedes explains part of the problem. Most
of the control methods which have been used before
now have relied on adding chemicals to potential
breeding sites to kill mosquito larvae, or involve spraying
or fogging with chemical pesticides which are designed
to kill off the adult mosquitoes. The image below shows

As well as being unpleasant for people, chemical

fogging in progress.

pesticides can also be damaging to the environment and

disrupt natural habitats. One of the biggest problems

Because Aedes aegypti live inside peoples homes

with these chemicals is that they dont just harm

remember all those empty bottles, tin cans, vases and

mosquitoes: other insects will also be killed (because of

bowls they breed in chemical fogging like this actually

this, we sometimes say that pesticides are

requires the pesticide to penetrate every room of a

indiscriminate).

house. You can probably imagine just from looking at the

picture below that this is not a very pleasant process, so

As a result, pesticide use can damage the ecology of an

people are often understandably reluctant to open their

area, reducing populations of insects which may be

doors and windows to let the chemicals into their house.

important food sources for birds and fish, or which are

As Haedes suggests, they may not always realise just how

pollinators for local plants and flowers.

close the mosquitoes are living.

Haedes also mentions another problem with this
approach: resistance. Pesticides rarely, if ever, kill all the
mosquitoes in an area there will be some which are
naturally resistant to the chemicals used. Because these
mosquitoes survive they will be able to pass on their
resistance genes to their children, so in the next
generation more mosquitoes will be resistant. Over time,
the proportion of resistant mosquitoes will increase in
this way, until eventually all or most insects in a
population may be resistant. As Haedes explains, many
Aedes aegypti populations are now resistant to the
majority of commonly-used chemical pesticides.

Because of all these problems, a

new approach to controlling Aedes
aegypti is needed.

Using genes to
control insects:
the Oxitec solution
Weve heard how traditional methods to control Aedes aegypti can be harmful to people and the environment, and arent
very effective in any case. In the film, the reporter then asks Haedes and Aegypta about the Oxitec approach. This is a
new technology based on advanced genetic science, which uses the natural instincts of the mosquitoes themselves to
track down other mosquitoes and stop them reproducing.
Before taking a look at the Oxitec solution in more detail, you can read a bit more about genes and genetic modification;
an incredibly powerful science which has already been central to recent advances in medicine, agriculture and
understanding diseases.

What exactly is a gene, and

what is meant by genetics?
Most people have heard of genes even if its not always quite clear what
they are, and what they do! A gene is the name given to a section of DNA.
In both people and mosquitoes, genes produce proteins that influence
everything from height to eye color. Offspring inherit these genes from
their parents
Genetics is the name given to the science of studying and manipulating
genes, and understanding how they are passed on from parent to offspring.
Over the last 50 years, since the discovery of the structure of DNA, genetics has
contributed to many important scientific developments, such as understanding
and treating inherited diseases like cystic fibrosis; developing more effective
targeted cancer medicines, like Herceptin; or enabling us to manufacture
large amounts of human insulin to treat diabetics.
Using modern genetics, scientists now have the ability to manipulate and
combine genes in many different ways. This allows them to study how genes
work, as well as combining different genes in ways that can be enormously
useful in medicine, agriculture and other applications.

Injecting DNA: Genetic modification

allows scientists to add to or alter
genes within an organism

Genetic modification (GM), also known as genetic

a chemical factory, synthesizing the human protein

engineering, is a biotechnology technique which can be

exactly. This has provided a quick and easy way to

used to add to or alter the genes within an organism. A

produce pure insulin for diabetics.

genetically modified organism, or GMO, is an organism

that has had its existing genes altered, or new genes

Since the 1980s, there have been a number of other

added, through a process of genetic modification.

human proteins produced through genetic engineering,

such as growth hormone or blood clotting factors.

Genetically modified organisms were first developed in

Genetic modification has also been used in a wide

the late 1970s and since then their use in a range of

variety of other medical applications: the Hepatitis B and

industries has become widespread. Perhaps without

HPV vaccines have been developed using genetic

appreciating it many people today are reliant on this

modification, and there is potential for scientists to

biotechnology for medicines and foods.

develop an entirely new field of cancer vaccines based

on this biotechnology.

One of the first products of genetic modification was

insulin produced by genetically modified bacteria.

Oxitec has applied the science of genetic modification to

Through insertion of the human gene for insulin into the

the problem of controlling populations of Aedes aegypti.

bacterias DNA, the bacteria effectively acts as

Applying genetic modification to insect control:

The Oxitec solution
As weve seen earlier in the video, the challenge we face in trying to control Aedes aegypti is to find ways of targeting the
mosquitoes where they live; in peoples houses and gardens. Whats more, we need to have ways of doing this which dont
harm people or other animals and plants in the environment, and which avoid the use of chemicals which mosquitoes are
often resistant to. One potential approach which meets some of these goals is known as the Sterile Insect Technique or
SIT.

Using sterile insects for

population control
The concept of SIT was first developed in the 1950s. The
basic technique is to dose male insects with radiation,
which makes them sterile. By sterile, we mean that
although the males do produce sperm and can fertilise

SIT uses the natural instincts of the released male

the females eggs, their offspring are inviable meaning

mosquitoes to seek out females, so it is much more

that they die at a very early stage of development.

effective than traditional means at targeting

difficult-to-reach pest populations, like Aedes aegypti. It

The sterile males are then released into the

is also species-specific: it affects only the target pest, and

environment, where they mate with wild females.

doesnt harm other insects.

Females usually only mate once, so a female which

mates with a sterile male doesnt produce any offspring.

Unfortunately, using radiation to produce sterile insects

As a result, the population as a whole is reduced.

in this way can cause problems. Not surprisingly, being

Eventually, with enough sterile releases, the population

hit by a large amount of radiation isnt very good for the

of the target insect in an area can be dramatically

male insects! Often, irradiated males are very sickly, so

reduced or even eliminated.

wild females prefer not to mate with them. If that

happens, they wont be very effective at controlling the

The Sterile Insect Technique was successfully used to

population. While SIT has been used successfully against

eradicate screw-worm (a pest of cattle) in North

some insect pests, mosquitoes are easily damaged by the

America. It has also been successful in reducing

process of irradiation, and to date there have not been

populations of other pests, such as eradicating the Tsetse

any successful programmes of mosquito population

fly, which causes sleeping sickness, in Zanzibar.

control using radiation-based SIT.

The Oxitec approach: genetic engineering of

sterile insects
Oxitec has developed a new way to control mosquitoes

Our modified mosquitoes produce high levels of this

using genetic modification. Our approach is similar to

protein because it actually activates its own gene,

the sterile insect technique, but because we use genetics

producing lots more of itself. Although its not toxic itself,

to stop our insects from reproducing, we eliminate the

it ties up some of the cells essential machinery. It can

need for damaging irradiation.

interact with other proteins which are needed for

controlling genes in the cell, and in this way it stops the

Scientists at Oxitec have developed a way to modify

cell from turning on other genes which are essential for

mosquitoes by adding a gene which produces a protein

it to survive.

that stops their cells from functioning normally.

All this means that the modified mosquitoes become
The gene produces a protein called tTA, which is a
special kind of protein able to act as a switch that
controls the activity of other genes.

very sick, and die before reaching adulthood.

How can we breed genetically sterile mosquitoes?

If the gene in the modified mosquitoes kills them, how does that make them
sterile? That depends on another special property of the gene, and the tTA
protein it makes: when the mosquitoes are reared in the presence of
tetracycline, it stops the tTA from working - in effect, it acts like an antidote.
So when we feed the modified mosquitoes with this supplement in the lab,
they stay perfectly healthy. But when the male mosquitoes mate with females
in the wild, their children inherit the lethal gene. Tetracycline is not present in
the environment in sufficient quantities to allow survival, so without the
antidote in their diet, the children of the modified mosquitoes die.
Because of this, the Oxitec genetically modified mosquitoes are effectively
sterile. When radiation is used to sterilise insects, as we saw earlier, their
offspring die at a very early stage of development before hatching. With the
Oxitec technique, the insects offspring die later in life, but the effect is the
same: when a genetically modified male mates with a wild female, her
children will die before reaching adulthood, so the population is reduced.

Why genetic modification works

So why use genetics? As we saw earlier, a major problem
with using irradiation is that the released males are
often sick, so females may not choose to mate with them.
In the video, Aegypta at first seems convinced that shell
be able to recognise Oxitecs males and avoid them. With
irradiated males, she might - but thats the clever part of
using genetic modification; because Oxitecs insects
dont have to be irradiated, they are fit and healthy. Like
all male mosquitoes, they will naturally seek out females
and mate with them. This means that Oxitecs approach
will be much more effective than other treatments, like
pesticides, at targeting mosquitoes in difficult-to-reach
places peoples homes and gardens. And unfortunately
for Aegypta and her friends, they wont be able to tell the
difference until its too late

No wonder Aegypta looks worried!

More on the science: How does Oxitec make

genetically modified insects?
To make a genetically modified mosquito, Oxitecs

After being injected, the eggs are hatched, and the

scientists have to find a way of incorporating the new

resulting mosquitoes carefully looked after until they

gene into the mosquitos own DNA, from where it will be

reach adulthood. Then they are bred with other

copied into every cell of the mosquitos body.

mosquitoes, and if the injected DNA has entered sperm

or egg cells, then it will be passed on to their offspring.

The process begins with mosquito eggs. These are tiny,

The DNA which was injected contains the lethal gene,

cigar-shaped objects about 1mm long. Using special

but it also contains a fluorescent gene which allows the

glass needles, so sharp that the point can only be seen

genetically modified mosquitoes to be identified using a

clearly under a high-powered microscope, Oxitecs

special microscope. So Oxitecs scientists can look at the

scientists can inject very small amounts of DNA into the

offspring of the mosquitoes which were injected to

end of a mosquito egg. The amount of DNA injected into

identify those which contain the new DNA.

each egg is miniscule thousands of times smaller than

a typical raindrop!

The scientists may have to inject thousands of mosquito

eggs to obtain just one individual which has the new

Many of the eggs injected in this way wont survive. In

DNA incorporated into their genome. But from this single

others, the DNA which is injected wont be incorporated

insect, a new strain of genetically modified mosquitoes

into the mosquitos cells. But in a very few eggs, the new

can be made.

DNA will be taken up by the mosquitos cells and will be

cut and pasted into the mosquitos own genome. If this
happens in the sperm cells of a male mosquito, or the
egg-producing cells of a female, the new DNA can be
passed on to their offspring.