Molecular Biology Genome Editing, Synthetic Biology, & the Ethics of Modern Science

Hello, and welcome back to www.educator.com, thank you for joining me.0000

This is going to be our last lesson in this molecular biology class.0003

What I wanted to do is talk about a few subjects that seem to be kind of glazed over,0007

if it even touched upon it all in normal college classes because they often run at of time.0017

There is certain information that needs to be covered.0024

While, this is very interesting stuff, it is not of the utmost importance.0027

It is really hard to lecture on.0032

In no way am I going deep into this, I’m just giving a surface level view and try just to get your mind working.0035

Try to get the topics in your head and just have you thinking about it a little bit.0042

We are going to talk about genome editing, synthetic biology, and stem cell biology.0048

We are going to talk about the ethical aspects of each of these, as well as just ethics of science in general.0053

Many of you who end up going to graduate school will actually have to take seminars in research ethics.0061

This is something that maybe will just be a starting point for you guys.0069

For any of you who work in a lab, this is going to be something that you always have to keep in mind.0074

It is always something that is going to surround your projects.0080

It is good, we need to think about this.0084

I’m going to do my job of introducing it to you and hopefully you take this upon yourselves to really think about these aspects.0088

Genome editing, let us start with that.0099

First of all, what is it?0101

It is genetic engineering in which DNA is either inserted, replaced,0103

or removed off from a genome using artificially engineered nucleases.0110

Remember, nucleases are the enzymes that cut up nucleic acids, DNA and RNA.0116

How do they work?0123

Double strand breaks get introduced by the action of the nucleases, at desired locations in the genome.0124

These double strand breaks then just get repaired by the normal mechanism0132

that we have talked about in this molecular biology class of homologous recombination or of non-homologous end joining.0136

Therefore, families of engineered nucleases that are in use for these processes today.0145

We have zinc finger nucleases, we have TALENs, we have meganucleases,0152

we have the CRISPR-Cas9 system or just the CRISPR system.0157

Meganucleases, they usually cut on average once per genome.0160

I'm going to focus on the CRISPR-Cas9 system because this is actually a really interesting system0167

and really cool things can be done with this system.0177

For genome editing, I’m just going to focus on this one.0180

Here is our example, here is our CRISPR-Cas9 system.0184

What is CRISPR? It stands for Clustered Regularly Interspaced Short Palindromic Repeats.0189

Cas9, the Cas proteins, that is CRISPR associated protein.0231

In this case Cas9, that is CRISPR associated protein 9.0250

What is this, basically what CRISPR is a very, let us say primitive, adaptive, immune system for bacteria.0255

If we are looking here, this is some membrane of bacteria.0280

It is their way of defending against a bacteriophage which is a virus that infects bacteria.0283

This is a primitive, adaptive, immune system for our prokaryotes, a bacteria.0295

This CRISPR, it is Clustered Regularly Interspaced Short Palindromic Repeats.0302

What it does is you have repetitions of base sequences in here, followed by spacer DNA such as this.0312

These are spacer DNAs.0330

Those spacers will recognize and cut, act like an endonuclease, cut up invading viral DNA.0334

It is almost similar to RNA interference but it is a different system for doing it.0346

How does this work?0353

First, the virus latches onto the bacteria, injects its DNA.0354

We have a double stranded viral DNA.0363

It can be simplified down to a 4 step process.0367

The invading viral DNA, double stranded DNA, or if it is an RNA virus, same thing can happen.0371

It is cut into small fragments.0386

From those fragments, you end up creating a novel spacer.0391

This is in DNA sequence and this is basically a way that this is linked to this specific piece of DNA.0397

The invading DNA or RNA is cut into small fragments, that is step 1.0407

Step 2, a copy of that invading DNA or RNA is integrated into the CRISPR locus as a spacer.0414

This one, this corresponded to a different piece of viral DNA.0426

This one to a different one and to a different one, and to a different one.0435

It is adaptive meaning it has seen that before, it keeps a piece of it so if it ever sees that DNA again,0441

it will be able to break that down and silence it.0448

Step 2, you make a copy of it, you put into that CRISPR locus as a spacer.0452

3rd step, the CRISPR RNA which is called a crRNA, CRISPR RNA, they get transcribed.0459

We will call this, this is step 1, this is step 2, this is step 3.0470

The CRISPR RNAs are transcribed from the CRISPR locus.0485

And then step 4, we have the crRNAs, the CRISPR RNAs, being incorporated into the CRISPR-Cas complex.0488

And then 5, it will target viral DNA, invading viral DNA and it will degrade that.0501

If want to call that, this is 6.0513

The crRNA will provide specificity.0516

It will act only, the crRNA, produced from these specific spacers, will specifically act on what is already seen.0520

Researchers can utilize this as the crRNA, the CRISPR RNA, providing specificity for such things as,0532

let us say gene editing for medical purposes.0543

You can silence something, let us say if you have an overactive gene that is causing the disease,0546

you can use the CRISPR-Cas9 complex to utilize that, you could silence that.0552

There is plenty more in depth detail to go into this that I'm not going to go into.0563

But I recommend you try to look this up, this is really cool, fairly cutting edge techniques being used here.0567

Gene therapy, specifically that is our delivery in nucleic acids into a patient's cell for treatment of a disease.0579

You can either be adding a gene, you can be knocking out a gene.0587

You could be replacing a gene, any of that.0593

The polymers of the nucleic acids are either expressed as proteins.0598

Therefore, interfere with protein expression or directly correct genetic mutations by replacing the actual DNA sequence.0604

The DNA that encodes a functional therapeutic gene will replace the mutated gene.0614

Therefore, when it is transcribed and translated, you will get the wild type protein,0621

instead of the mutant protein that has been causing the disease.0626

Gene therapy, the theory on it was first published in 1972.0632

The first experiment approved by the FDA occurred in 1990, about 18 years later.0638

13 years after that, the first commercial gene therapy was approved at for use as a cancer treatment.0645

The gene therapy called genidicine was approved in China in 2003.0653

An example of a gene therapy, right here, we can see this is gene therapy using an adenovirus vector.0664

An adenovirus is a specific type of virus, what it can do is you can manipulate the DNA inside the virus.0672

The virus will infect the cell, bring that DNA into the nucleus of the cell.0680

That DNA now being a good piece of DNA, not something we need to worry about of like viral DNA.0686

But something that we plan on putting there, that will be healthy for the cell,0693

will then make the certain proteins that we wanted to make and will help, if used properly to treat that disease.0697

This is a little different from RNAi or from the CRISPR system, in the fact that we have something0707

that we always need taking into consideration, when we involve a virus.0714

Yes, it is there are plenty of research on it and plenty of ways that it has been altered,0720

so that it should not be able to replicate and cause disease as any normal virus would.0729

As always, something as a scientist and as the public, you keep in the back your mind.0735

But for the most part, it does its job without infecting quote and quote, in a bad way, but infecting in a good way,0740

getting in the cell, giving its DNA which has the good DNA that we want to make the protein of interest.0748

Some ethical questions revolving around gene therapy is, there is always going to be problems of genetic engineering.0758

Maybe the protein that you wanted to be sent in there.0766

Maybe there is a mutation on the gene.0771

Maybe you did not engineer the gene properly.0773

Or just maybe the ethical question of just revolving around genetic engineering itself,0776

manipulating the genome, playing god, if you want to say it that way.0784

Another question that always is going to surround gene therapy is the aspect of gene doping,0790

meaning, basically, if used improperly, you could almost use it as a performance enhancing drug alteRNAtive.0797

You can do gene therapy to make you bigger, faster, stronger, have more oxygen carrying capacity, something like that.0810

That is always a question that will be thrown around.0819

Moving on to a different topic, synthetic biology.0827

Synthetic biology is described as the design and manufacture of biological components that do not exist in nature.0831

There are many ways of look at synthetic biology.0840

I want to look at a specific one.0843

This is an example of synthetic biology work.0847

In 2010, scientist at the J. Craig Venter institute, if you remember J. Craig Venter,0852

he was the entrepreneur that was involved in pushing the private funded aspect of the human genome project.0856

Scientists at this institute had been opened, run by him, created the first synthetic cell.0869

What they did is they started with a bacteria called mycoplasma capricolum.0876

What they did is they took out all of the DNA, any genetic material in there,0882

and replaced it by using the transformation technique with a brand new fully synthesized,0889

fully synthetic, fully brand new DNA, made from their typing on a keyboard.0898

They made it in a computer, made a bunch of DNA, you can get DNA made.0907

They inserted their computer coded DNA into this completely empty cell.0914

It is like a shell at this point.0922

They obviously tried it many times until one work.0924

And they finally found a sequence that was actually able to replicate the cell, as if it were a normal cell.0926

In this case, they actually created a new species that they called mycoplasma mycoides.0937

This is huge, they were able to take a manmade coded DNA sequence and put it in a shell of a bacteria, to make a living replicable organism.0944

This is a huge, absolutely huge event.0964

That was such a huge event like that, there is always going to be ethical questions.0971

One of the big questions that surrounded this was, did they create new life or did they just manipulate life?0978

You can have your own opinion on this, one of the rulings I believe it went to the court system was that,0992

this was not actually the creation of a new life.0999

However, this is something that will always be asked about.1002

Some other ethical questions regarding synthetic biology is genome always are an ethical debate in and of itself.1009

But when you start being able to computer code genes and be able put it into an organism,1016

that brings up an entire new set of questions about gene.1024

Finally, the fear of these synthetic bacteria that they made does not exist in nature.1030

It was not intended to exist in nature.1039

We have made it exist, we as a human race made it exist.1041

What is that going to, what type of effect will that have on the current ecosystem, the currently living system?1046

Can that bacteria infect other species?1055

They took very good precautions to make it a non infective bacteria, in non-virulent strain.1059

But there are ways that it can mutate or there are ways that,1066

maybe somebody else might not take as great of care or might use this for ill purposes.1070

That is always going to be an ethics debate.1078

Moving on to stem cell biology, I know this is a big hot topic.1082

This is been ethically debated, since the thoughts of stem cell biology came out.1086

What is stem cell biology or using stem cells to treat or prevent disease.1093

Often you have your stem cells coming from the bone marrow.1097

When you talk about bone marrow transplant, a lot of times people with cancer,1104

or multiple myeloma, they are one of the specific ones.1109

You need to get bone marrow transplant and that is because cancer,1113

as well as the chemotherapy, ravages your quickly dividing cells such as your B and T cells.1117

Your B cells, part of your immunity that is found in your bone marrow.1124

You need bone marrow replacement.1128

You have what are called hematopoietic stem cells, these are stem cells that can turn into any type of white or red blood cell.1131

Your B and T cells are a type of white blood cell.1139

This is very useful and a very common practice nowadays.1143

Some people do not agree with the fact of umbilical cord blood being utilized as a source of stem cells.1149

In a huge contest or point of contention, is the fact of actually using embryonic stem cells.1157

Stem cells from fetuses or unborn fetuses, that is a huge point of contention1166

with many groups especially your religious focus groups.1173

As well as even some nonreligious focus, some scientists do not agree with it.1179

You can also have embryonic stem cells.1184

You can also have what is called somatic cell nuclear transfer technique.1186

But you need to start with some sort of a cell to begin with.1190

The good thing about stem cell biology is that it can be used to treat many types of diseases.1196

You have leukemia lymphoma, diabetes, injuries to the brain and spinal cord, deafness, anemia.1204

Anemia is the low concentration of red blood cells which will then affect your oxygen carrying capacity.1214

As well as stem cell biology can be used to treat organelle transplantation, to hope that goes well.1225

Some of the ethical questions, are you destroying life?1234

If we are talking about using embryonic stem cells, are you actually destroying a life that should have been?1239

That is going to be a point of contention that people are going to have, a side versus the other.1247

A lot of these decisions on ethics, you try to use as much as possible.1254

But really, it is going to come down to belief systems and1260

how you can manage that between yourself and the people that you have a contention with.1263

Another one is human cloning.1270

If we are using stem cells to make other humans, that is a huge point of contention.1272

As of right now, human cloning is not allowed.1281

Will it be in the future? Maybe, we do not know.1286

Other than the ones that I had mentioned before, there are many other topics that are up for ethical debate.1292

Some that I wanted to list here, genetically modified organisms.1299

Vaccines, the whole vaccines, people will be thinking that they are correlated to autism.1304

Based on a study published by a medical doctor who named Andrew Wakefield, in The Lancet,1313

who was actually paid by competing vaccine company to falsify data1323

and talk bad about the vaccines that are out on the market, which has spawned even though he lost his medical license.1330

Once it was found that he falsified his data, the paper was retracted but a lot of people do not hear that part of it.1339

They only hear the fact that the story was published years ago.1346

This is always going to be for debate.1351

A lot of people do not have the right facts to follow that one specifically.1353

We have other topics for debate like save your babies.1358

A save your baby is when a couple has a child with a condition where they are dying because they lack,1361

let us say for example they have a kidney disease and they need a kidney transplant.1374

Sometimes a couple will have another baby that was basically meant1379

to use that second baby as basically an organ farm for the treatment of the first child.1385

The second baby was born so that it can give a kidney transplant to the first child who is sick.1395

There is always going to be ethical debate over that.1406

There is ethical debate on genetic manipulation.1410

Gene editing for health, you will have people on both sides.1413

As well as the build –a-bear type baby, I want to make my baby blue eyes, 6’5, highly athletic and highly intelligent.1419

Who is to stop somebody from picking all the best qualities to make their child, just like a freak of nature?1433

I want to make somebody with the basketball skills and body type of Le Bron James, with the mind of Albert Einstein.1443

It is something crazy like that.1454

Prenatal genome sequencing, for the same reason, on being able to do that build-a-bear baby.1458

Genome sequencing and insurance companies.1464

If I sequence my genome because I want to know what diseases I’m susceptible for.1467

Insurance companies are allowed to have that data, they might not offer me insurance at all.1474

Or might have an extremely high premiums, based on my risk factors.1482

That is a huge debate.1488

Cloning, always a big one, specially human cloning.1490

Animal testing, big debate.1493

You are always going to have groups that PETA and even other scientists telling you that animal testing1495

does not need to happen or does need happen.1503

You do need to test on animals, if you want to pass selected drugs through the FDA, to get to the market.1507

Like your Advil, your Tylenol, even all the way up through your chemotherapies,1518

they have all had to have been tested on animals, before they allowed to go in the clinical trials and be tested on humans.1525

Even though there is ethical debate on this, there is a specific way that you have to follow,1534

to be able to get your drugs on the market.1542

And then, one that is a different type of ethical debate is falsifying data.1547

As a scientist or as a future scientists, this is extremely important.1558

Falsifying data, whether it is for any reason, sometimes people falsify1565

because it will help them get a promotion or maybe they work for a drug company and they are trying to push their product.1571

Maybe they just want to become famous, whatever the reason.1578

This has a huge detriment to the scientific community.1581

Not only does it make people in the scientific community and outside the scientific community distrust the overall scientific community.1586

But it can actually hurt the current focus of research because if someone comes out and says that1598

this is an answer to a question that everyone has been looking for, and they falsified all your data,1612

you do not have a lot of people able to back up their results but maybe just believe it.1621

Then, you take that for granted that that is the correct answer.1628

It can set you back 50 years.1633

It can end up causing you to go down the rabbit hole.1635

This is a huge deal that everyone needs to have high morals, when creating their research.1639

They need to give their data, not falsified data.1647

Do not cherry pick your results, you give all your results not just the ones that make your story look good.1652

If you do the experiment 10 times and 9 out of 10 times it fails, but one fails does not gives you an answer you do not want.1661

But the one time, it gives you the answer that you do want, that is the only piece of data that you share with the public, that is a problem.1670

That is a big ethical problem.1677

Let us talk about research ethics in general.1684

Research ethics involves the application of fundamental ethical research principles to variety of topics involving research.1687

Examples will include the design and implementation of research, involving both animal and human experimentation.1700

As well as scientific misconduct, like the falsifying data falls under that.1706

The declaration of Helsinki which is set forth in 1964 is a set of ethical principles,1713

regarding human experimentation developed for the medical community by the WMA, the World Medical Association.1724

This is the first fairly complete document that most countries adopted.1731

The basic principles of this declaration of Helsinki was that, of utmost importance is the respect for the patient.1739

Their right to self determination and their right to make an informed decision which we hear very often,1749

informed consent, we hear that in hospitals a lot in the US.1756

It is important that they know what they are getting themselves into in a certain research, trial, let us say.1763

What is important is at the researchers duty is solely to the patient or volunteer,1772

meaning they care most importantly about the patient, overall, even over the research.1778

As hard as that is to think about, as a researcher, the patient is the most important part.1788

Patient welfare takes precedence over both scientific and societal interests.1796

Ethical considerations will take precedence over laws and regulations.1804

Very importantly, same thing that we see in the hospital system as well.1810

If a research participant is incompetent, physically and mentally incapable of giving a consent or is a minor,1816

then you need to get surrogate consent from somebody else.1823

The last few slides, I just want to talk about what is right vs. wrong.1833

When we talk about ethics, you talk about right vs. wrong.1841

The ultimate debate, ethics is the ultimate debate of right vs. wrong.1846

There is no correct answer, there are maybe one that you favor but there is no truly correct answer, when we are talking about this.1852

The question is going to be asked forever.1862

Much of modern biotech has a markedly lifespan and health span focused approach.1864

Lifespan, making people live from saying not just 80 years old to 100 years old.1871

Health span is actually being healthy until you are 80 instead of 70, something like that.1878

A lot of that is a morally, good morally, just goal.1888

There are still lingering fears that exists that scientists have too much control over human evolution and destiny.1894

Because the better we get with technology and biotechnology specifically,1902

the more control we have over the human body, the animals, plants, and bacteria around us.1906

It is that fear of we are becoming a godlike figure, we can affect destiny.1917

In additional laws and regulations, both public and political opinions1928

can affect the current and future state of research and development.1933

This is something that we are forever going to have to work with, that is not going away.1936

Scientists cannot just stay in a hole and do their research, without worrying or taking into consideration1945

what politicians and the public have to say about it, for better or worse.1953

Let us talk about a specific aspect.1963

We will bring up Dolly the sheep, if any of us remember that.1966

This happened in 1996 in Scotland.1969

Dolly was the first animal to be cloned from an adult’s somatic cell, using the process of nuclear transfer.1972

What that was is, the nucleus from an adult cell is transferred into an unfertilized, enucleated, without a nucleus zygote, which is an egg cell.1980

You have an egg cell, you take out the nucleus.1995

You take the nucleus from an adult cell, from a different animal.1998

Take the nucleus out, put it into the egg.2004

Zygote enucleated, adult cell with the nucleus.2015

Take that out, put it into here, and then try to grow an organism.2020

They actually did this with the sheep.2028

That sheep lasted about 6 years, before passing away.2031

This brought up a lot of questions and will continue to bring up a lot of questions.2037

The fact of, is science unstoppable or can we actually limit how far we go, based on rational ethical thinking?2041

Just because certain science is possible, does not mean that it will actually come to fruition,2054

if we as scientists or researchers think rationally and ethically.2060

But the public always fears that science and scientist would not.2066

It will always be, we want to do this just because we can do it not because we think it is going to be beneficial.2072

That is why this is kind of, just because we can, should we, type of the question.2080

What is going to happen is that, it may fall upon the scientific community to start the ethical discussion within itself.2088

As human biotech research progresses, as it is doing right now,2097

ethical debates will continue to heat up and will continue to be extremely hot.2101

Not only actions but also the intentions of the scientific community require rational reasoning and justification.2108

An example here for ethics is the whole genome sequencing.2120

Pretty expensive in general but it is becoming cheaper and cheaper.2126

We have that $1000 genome insight, meaning, I might, in the near future be able to send in a sample of my DNA2130

and for only $1000 get my entire genetic profile, which would be amazing.2138

But whole genome sequencing differs from current prenatal genetic testing.2147

First of all, current tests identify only serious genetic conditions in fetuses, at about moderate to high risk.2153

Whereas prenatal whole genome sequencing could allow for the entire genetic profile.2165

This could lead to the possibility of wanting to make that build-a-bear type kid.2173

I can see that my kid will have brown eyes.2185

Maybe we can use some gene therapy and give him or her, blue eyes.2188

Or maybe, my child is going to be a boy, is it early enough to where I can change it to be a girl?2193

Or my child will have autism or another type of disease that is not considered seriously fatal, that is not currently being tested.2201

I think we only test for somewhere, I could be wrong, I think around 13 to 15 different genetic conditions.2221

But this can lead to a situation where me and my child will have chondroplasia, might be a little person.2229

Any of these things that are not perfectly in line with what the parents wanted,2240

they can either fix that to their liking or decide to not proceed with the pregnancy.2248

A lot of these ethical issues will come up.2256

Also, this whole genome sequencing, as I said before, you can get your whole genome profile2259

but maybe the insurance companies, the health Insurance companies get a hold for that.2265

They would be able to possibly then decline you, based on your risk of a certain disease, big ethics problem.2269

On the good side, golden rice, a genetically modified organism.2283

This is a rice strain that was modified to contain β carotene.2288

β carotene is a precursor of vitamin A.2293

A single serving of this golden rice supplies 60% of the recommended dietary allowance of vitamin A.2297

This is important because according to the World Health Organization,2308

more than 215,000,000 preschool age children are vitamin A deficient.2313

Over 2 million deaths in more than 500,000 cases of blindness are due to vitamin A deficiency worldwide.2321

Just the introduction of this strain of rice can help decrease this.2332

In fact, there have been estimates that over a million people have been saved from dying,2340

in the several years that the golden rice has been on the market.2351

It will just continue to rise, as year by year goes by.2359

This is a good aspect, however, still going to be ethical debates.2364

There is always going to be people who are for or against genetically modified organisms.2367

I had just last two slides and I want to leave you with some questions to ponder.2377

When thinking about research, any specific research, you want to know what is the societal impact?2383

Are any human rights being violated?2391

That is a big one.2393

Do the benefits outweigh the risks?2396

Do any means justify a good end?2400

The means justify the end, right?2406

Or sorry, does the end justify the means?2408

But does any means, how you get to that result, is that okay?2410

Does biotech ultimately advance or impede the common good of humanity, and the world as a whole?2416

This is probably the toughest one to think about, is there anything that we have not thought about yet?2426

That is a tough one to think about because you think about everything, and then there is always something later.2432

This one you really have to prepare, prepare and think.2439

How do you answer questions on ethical debates?2454

The biggest thing that I can tell you is doing your own research, do your own literature research.2461

You should be the expert on all the experiments that have been done before, both for and against.2471

You want to know both sides of the argument, why it is an ethical debate, why it is bad.2482

You need to do your literature research, that is a huge thing.2494

You need to understand the science and you need to understand the ethical ramifications from this.2497

This is extremely difficult, even for people in the scientific community, literature and analysis is a tough job.2504

It is tough to do, you get better every time you do it but is really tough.2517

It is a tough skill to master.2524

For those of you outside of the scientific community, it is even harder because2527

they are not strong in the subjects of biology, of genetics, of biotechnology.2532

They do not understand it, they might not understand why you would use a certain experiment2538

or why you might have to use animal experimentation.2544

A lot of the disagreements come from the fact that many people are uninformed or many people do not understand the science.2550

The media, the scientists are not usually the ones who is talking straight to the public,2561

the media are the ones talking to the public and the politicians.2566

The media, as much as we want them to know everything,2570

they cannot be expected to understand published scientific data on their own.2578

That is going to land on the scientific community.2584

We need to educate the general public and especially those in the media who cover scientific topics.2589

It falls on our shoulders, we need to educate the media, we need to educate the public2597

so we can bring them up onto the knowledge level to understand much easier.2602

But once we educate the public, especially once we educate the media,2610

the media is responsible for taking it upon themselves, taking it upon themselves to gauge their level of understanding,2616

before writing articles that will reach the masses and possibly promote fear mongering.2631

The media needs to understand the science, before writing an article that could cause a huge problem in the nonscientific community.2640

We, as scientists, can only do our best to educate the community, to educate the media.2656

The media has to take it upon themselves that if they do not really understand what is going on,2662

they need to reach out and do their own homework and get in touch with the scientists to prepare themselves.2668

That is the end of the lecture, the end of the course.2676

I wanted to say from myself, thank you so much for joining the course.2680

I really hope you enjoyed it, and from www.educator.com, we appreciate you seeing this course.2685

Please check out some other courses, there are plenty, from high school through the college level.2692

From me, and www.educator.com, thanks for joining and we hope to see you again.2700