PROJECT O.E.C. Angela Liu O IL E ATING C RUSTACEANS.

PROJECT O.E.C.

A n g e l a L i u

OIL EATING CRUSTACEANS

Where I live

BP Oil Spill Area

TEXAS

INTRODUCTION• BP oil spill (April 20, 2010)– Gulf of Mexico– 4.2 million barrels of oil – One of the worst environmental disasters in

U.S • Direct result?– thousands of native species injured/killed

• 5 years later?– Millions of gallons of toxic oil remain on sea

floor

PURPOSE

• removing oil left on seafloor

• soften the impact of future oil spills

• How?– genes of hydrocarbon-

chewing microbes + germ cells of squat lobsters

– CRISPR-Cas9 gene drive method

Methylococcus capsulatus

C O M P E T I N G T E C H N O LO G I E SMethod What Pro Con

Dispersants

chemicals sprayed on surface oil that break oil down into smaller droplets

quick Harmful chemicals

In situ burning

the remaining surface oil is burned

quickly burns large volumes

Pollutes air

Oil-eating microbes

microorganisms that are genetically engineered to break down oil

not bad for the environment

-cannot clean the deep seafloor because it’s too cold - microbial predators like protozoa exist

Booms floating barriers blocking oil from spreading

can prevent oil from spreading

- can only prevent oil from spreading- only useful in calm water

Skimmers

devices that remove oil from the surface of water

very easy to operate and will collect almost all forms of oil

- only work in calm water- Plastic and other trash often clog them up

the DESIGN

• Methylococcus capsulatus– an immobile, coccoid (spherical) bacterium

that has methane monooxygenases (MMO)• MMO are enzymes involved in the

bioremediation of oil–Oxidize the hydrocarbon bonds found in

methane• chemical formula: – CH4 + O2 + NAD(P)H + H+ -> CH3OH +

NAD(P)+ + H2O

MMO FORMULA breakdown

• the process by which the enzyme MMO oxidizes methane into methanol, breaking down the hydrocarbons found in oil

the DESIGN (cont ’d )

Inserting MMO into Squat Lobsters (chirostylidae)• CRISPR-Cas9 method will be utilized– a genetic engineering mechanism

invented by researchers and scientists at the Wyss Institute for use in plant/animal cells

CRISPR-Cas9

• How does CRISPR-Cas9 work?– Cas9 is a naturally occurring enzyme that

scans for foreign DNA from bacteriophages (viruses) within the host genome’s DNA

– acts as a pair of molecular scissors -> cuts specific DNA sequences within the genome.

– the cell attempts to repair the cut itself..but the repairs are mutated

– a mutant gene sequence can be replaced with another piece of DNA with a more desirable sequence.

the DESIGN (cont ’d )

• MMO enzyme will be inserted into the fertilized egg cells of squat lobsters (specifically chirostylidae)

• their offspring will all be transgenic animals with beneficial oil-eating mutations

• squat lobsters are a wild species, it would be very hard to maintain an altered gene sequence

• Thankfully… the CRISPR-Cas9 gene drive method exists

t h e C R I S P R - C a s 9 G e n e D r i v e

• when the squat lobsters reproduce:1. The Cas9 enzyme from the gene

drive squat lobster cuts off a specific gene sequence from the wild squat lobster

2. The cell pastes a copy of a sequence from the altered chromosome into the break in the chromosome of the wild type gene

• the altered gene drive will be passed down through next few generations

GENE DRIVE in squat lobsters

EXPECTED RESULTS

MMO is present

Using CRISPR-Cas9 Gene Drive

Majority of squat lobster population is oil-eating

1 0 0

0 1 0

1 1 1

0 0 0

Truth Table (assuming design works perfectly)

ADVANTAGES

• Previous methods cannot reach the gulf’s sea floor, on which the majority of the oil has now settled

• oil on the seafloor can easily be accessed by the oil-eating squat lobsters, since the seafloor is the squat lobster’s natural habitat

• oil-eating microbes are eaten by microbial predators like protozoa, whereas squat lobsters can survive much longer

• highly effective - only one wild squat lobster will need to be genetically altered for the oil-eating properties to spread throughout the entire population (by using the CRISPR-Cas9 gene drive mechanism)

ENVIRONMENTALLY

ECONOMICALLY

POTENTIAL PROBS

• CRISPR-Cas9 system is recent -> many risks

• no way to fully test the effectiveness w/out actually releasing them into the wild

• unknown consequences of cutting a gene sequence– may delete a fxn the squat

lobster will eventually need• economic and environmental

rewards outweigh the potential problems

TESTING

1. Initially tested on a single squat lobster -> see its efficacy and oil-eating capabilities

2. Genetically modified squat lobster will be released into a controlled population of wild squat lobsters within an artificial environment replicating the one that squat lobsters actually live in

3. The squat lobsters’ living and reproduction will be carefully monitored for a couple months

4. Once the success of the genetically engineered squat lobsters has been confirmed through more than 5 generations, they will be placed into the Gulf of Mexico

WORKS C ITED• "At the Bottom of the Gulf of Mexico, Corals and Diversity Suffered After

Deepwater Horizon Oil Spill." NOAAs Response and Restoration Blog. N.p., 01 Apr. 2015. Web. 08 July 2015.

• Das, Nilanjana, and Preethy Chandran. "Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview." Biotechnology Research International. SAGE-Hindawi Access to Research, n.d. Web. 08 July 2015.

• "Deep-sea Crustaceans." NOAA Ocean Explorer Podcast RSS. N.p., n.d. Web. 08 July 2015.

• "Dispersants." Dispersants. N.p., n.d. Web. 08 July 2015.• "5 Years after the Gulf Oil Spill: What We Know - CNN.com." CNN. Cable

News Network, n.d. Web. 08 July 2015.• "Gulf Oil Spill: Still Poisoning Dolphins to Crickets." Student Science. N.p.,

n.d. Web. 08 July 2015.• "Gulf Oil Spill: Still Poisoning Dolphins to Crickets." Student Science. N.p.,

n.d. Web. 08 July 2015.• "Second Opinion Doctor." Second Opinion Doctor RSS. N.p., n.d. Web. 08

July 2015.• "Slick Solution: How Microbes Will Clean Up the Deepwater Horizon Oil

Spill." Scientific American Global RSS. N.p., n.d. Web. 08 July 2015.• "World Socialist Web Site." The Social and Economic Impact of the Gulf Oil

Spill -. N.p., n.d. Web. 08 July 2015.