Using Gene Editing To Repaint Butterfly Wings Mit Genmanipulation Schmetterlingsflügel neu streichen Edición genética para repintar las alas de las mariposas Usare l'editing genico per ridipingere le ali delle farfalle 유전자 편집으로 나비 날개를 다시 칠하기 Genbewerking gebruiken om vleugels van vlinders te verven Wykorzystanie edycji genów do przemalowania skrzydeł motyla Usar a edição de genes para pintar as asas das borboletas Использование редактирования генов для перекрашивания крыльев бабочек 利用基因编辑重新绘制蝴蝶翅膀 使用基因編輯重新繪製蝴蝶翅膀

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How… to paint…a butterfly wing.

Hey smart people, Joe here.

CRISPR: it's a DNA-editing technology that you've probably heard about in terms of

disease, medicine, maybe making genetically modified organisms.

But scientists are using it for some really interesting questions, like why do butterflies

have such awesome looking wing patterns, and how do they form?

So I'm here at George Washington University.

And I'm gonna go CRISPR some butterflies.

Now, there's been a lot of hype around CRISPR.

CRISPR RRRRRRRRRRRRR

But what is it actually?

CRISPR is a DNA hacking system with two parts.

One part is a piece of RNA that carries a set of coordinates matching a specific spot

in the genome's DNA. in het DNA van het genoom.

The other part is a protein that chews through DNA, which creates a small mutation. Інша частина - це білок, який пережовує ДНК, створюючи невелику мутацію.

And we can program CRISPR with a specific set of coordinates so it cuts exactly what

we want.

AM: You see this red stuff here?

JH: Mhmm AM: This is CRISPR (wink)

JH: A tube full of CRISPR AM: CRISPRRRRR JH: Een tube vol CRISPR AM: CRISPRRRRR

JH: Ahhhhhh JH: So, everytime you hear someone say CRISPR,

now you know what it looks like.

That's Dr. Arnaud Martin.

Dr. Martin and his team are using CRISPR to understand how butterfly genes make so many

crazy patterns and colors.

There's more than 200,000 species of butterfly and moth – all with their own unique wing |||||||moth||||||

patterns.

We know they use those patterns to attract mates, hide from predators, and send warning

signals, but how and why these colors get painted is still a mystery.

But this is about more than just studying butterfly patterns.

These scientists are trying to answer an important question about our own biology and even life

itself: How do the instructions in DNA build bodies?

I mean, genes–the letters of DNA–are just codes.

How do we go from those letters and codes to the many beautiful shapes and colors we

see in nature?

This is a question CRISPR can help us answer.

AM: Those fundamental basic questions of how genes make shapes, this is relevant to us.

I mean, what I want to understand is how DNA makes, you know, people.

The first step to figuring out the mystery is easy: collect some butterfly eggs.

This is Joe.

This is also Joe.

He's a researcher in the lab.

So, we're on the roof of a building in downtown Washington DC, in a greenhouse.

JH: That's why I feel so tropical.

OJ: Yeah, it's maybe 72 fahrenheit in here.

Maybe a little warmer.

And about 85% humidity.

We keep Gulf fritillary butterflies here. We houden hier Golfparelmoervlinders.

If the team is lucky, they can collect around 40 eggs a day from these butterflies to modify

with CRISPR.

JH: These are one of my favorite butterflies.

They're super pretty.

They have these lovely silver patches on the underside of their wings, which I just think

are really, really beautiful.

JH: So you wait for the butterflies to lay enough of the eggs, and you collect them so

you can do the work you're going to do?

Exactly!

What we do with CRISPR, rather than being super precise

we're sort of going in with a hammer and smashing the gene and then seeing what happens.

It's like if you wanted to understand how a car worked, so you open the hood and just

started smashing pieces.

And then found the way in which the car stopped working.

If the car just completely stops, then maybe that doesn't tell you anything.

But if the car still works, except the radiator is now broken, then you understand that the

bit you smashed has something to do with the radiator.

So that's the version of this that we're doing.

Very broad strokes, breaking bits and seeing what breaks. Heel brede slagen, stukjes breken en kijken wat er breekt.

The next step is we take those eggs down to the lab to inject them with CRISPR.

And by we, I mean me.

I'm going to do it.

Alright, your turn!

Here we have a Gulf fritillary egg from the top |||||fritillary butterfly|||| Hier hebben we een Gulf fritillary ei van de top

You move the needle back, you approach gently, you get in, and you press the pedal.

There it is!

I did it!

Oh you can see the little red burst inside.

CRISPRRR

The eggs will develop and hatch like usual, only the DNA inside has been altered by the

CRISPR that we injected.

The caterpillars, look, well, like normal caterpillars.

You'd never know the difference.

Unless you look inside their bodies.

Okay, let's talk metamorphosis.

You've maybe heard that when a caterpillar morphs into its final form, inside the chrysalis,

it completely liquifies into soup, and that liquid rearranges to form a butterfly. het wordt volledig vloeibaar tot soep, en die vloeistof herschikt zich om een vlinder te vormen.

This misconception has been repeated so often it's replaced the truth.

And what actually happens is way cooler.

Caterpillars mature from the inside out.

The larvae move through stages of growth, called instars. De larven doorlopen stadia van groei, stadia genaamd.

When an instar gets big enough, it crawls out of its skin and the next stage of growth ||larval stage||||||||||||||| Wanneer een instar groot genoeg wordt, kruipt het uit zijn huid en de volgende groeifase

emerges from inside.

And when the caterpillar is just about big enough to form a chrysalis, it already has

some pieces of the adult butterfly inside it…

What you're about to see absolutely blew my mind: Wat je op het punt staat te zien, verbaasde me absoluut:

So, you see this and you're thinking no way this thing has wings, it's a larva,

it's not even flying!

What the heck…

I'm going to make an incision between the two nostrils, between the diaphragms. |||||cut|||||||

Check that out.

This is incredible.

That… is….

That's a larval wing. Dat is een larvenvleugel.

That's a baby wing!

Here we go!

You can see the veins and everything, it looks like a tiny, clear butterfly wing.

Wow!

That's right.

This is the stage where not only the shape of the wings is defined, but also the position

of patterns.

That's right, caterpillars have baby butterfly wings inside them.

And even at this early stage, the butterfly's wing pattern is being painted.

The team can label which genetic instructions are turned on in that baby wing.

And what's crazy is where we see certain genes turned on lines up perfectly with where

the patterns will be on the adult butterfly.

And when CRISPR messes up that DNA instruction?

We can also see how the pattern is disrupted.

So the different genes that you study here in the lab lay down different parts of

this pattern?

Exactly, so during larval development you have a canvas of cells that are communicating,

and the wings need to decide where to make, maybe, reflective scales, or dark scales. en de vleugels moeten beslissen waar ze reflecterende schubben of donkere schubben moeten maken.

And it's really, a little bit, – if I can make an analogy – of sketching process,

where the outlines of each patterns are determined super early.

It's during metamorphosis in the pupa or chrysalis that really the scales are emerging, Het is tijdens de metamorfose in de pop of pop dat echt de schubben tevoorschijn komen,

and the colors happen.

It's like a paint by numbers.

The genes they've identified draw in the boundaries and say “paint here.”

Later on, inside the chrysalis different genes paint in the colors based on those early instructions.

But the basic shapes, the organization, the concentric rings, stripes – the position |||||||nested circles||||

of all the system is established super early in the larva.

Which is mind blowing.

So now you know caterpillars don't turn into total mush as they mature, and they have

some adult body parts hidden inside them.

BUT!

There's still a ton we don't know about how wings form inside the chrysalis. Er is nog steeds een heleboel dat we niet weten over hoe vleugels zich in de pop vormen.

If only we could see inside.

Well some scientists have figured out a way to do that, like our old friend Aaron Pomerantz

a PhD candidate at UC Berkeley:

What my lab tries to understand is how butterflies form their wings and their scales, which occurs Wat mijn lab probeert te begrijpen, is hoe vlinders hun vleugels en hun schubben vormen, wat gebeurt

in the pupal stage. in het popstadium.

Now If you've ever stared a pupa for long enough, you may have been a bit underwhelmed. ||||||||||||||||disappointed or unimpressed Als je ooit lang genoeg naar een pop hebt gestaard, was je misschien een beetje teleurgesteld.

It doesn't look like they're doing a whole lot.

They don't really move. They don't often look that flashy. Ze bewegen niet echt. Ze zien er niet vaak zo flitsend uit.

But just below the surface there's an incredible amount of change happening.

Caterpillars do contain the precursors to their adult wings: a small cluster of cells

known as an imaginal disc. bekend als een imaginaire schijf.

And these cells have all the information necessary to transform into an adult wing when the time

is right.

A couple of scientists- Julian Kamura and Ryan Null–figured out on accident that if you remove this imaginal

disc, now you would have a window into the pupa.

So now we can set up a time lapse under a microscope to watch this entire process happen.

And what we see is *incredible*.

The cells in the immature wing start to specialize, or differentiate into elaborate shapes and colors

Those gene instructions, laid down in the baby butterfly wing, tucked inside the caterpillar, ||||||||||hidden away|||

tell the wing where to paint in these colors

It's both fascinating to me, and important to science, that we can watch the wings as

they develop, and see how colors are filled in.

The adult butterfly wing is covered in thousands and thousands of scales, and this is where

the color comes from, because each one of those scales produces a specific color - either

through the architecture of the scale that creates a certain wavelength, known as structural

color, or from pigments that become deposited inside those scales.

…in the CRISPR mutants, some of those cells are broken, like the car's radiator, so

we can see how that changes the wing pattern.

When metamorphosis is complete, the butterfly that emerges is called a mosaic mutant

It has a change in some part of its body.

Here is the butterflies we had in the cage over there, Agraulis, where you have Hier zijn de vlinders die we daar in de kooi hadden, Agraulis, waar heb je

these lovely precisely placed silver spots all over the wing surfaces.

And then we knock out one gene - a gene called WntA.

We literally just go in and smash it with a hammer so it's not there anymore, and

what we get is this.

There are still silver spots, but the arrangement of those silver spots is completely different.

In other butterfly species, switching off that gene had totally different results:

it can make patterns fade, or even disappear.

WntA seems to be the master sketching pencil for butterfly wing patterns.

And they've identified another gene, called optix, that's more of a master paintbrush.

Messing with it can turn some butterflies black, and make others iridescent blue.

These genes are part of the master set of instructions to build a body, and we have

similar genes in our bodies.

We can't go in and break those genes in humans to understand how they work, but we

can learn something about them by decoding how these beautiful insect patterns are painted.

When people talk about CRISPR they like to think of creating mutant creatures or superhumans, Als mensen over CRISPR praten, denken ze graag aan het creëren van gemuteerde wezens of supermensen,

but here in real life, CRISPR has given scientists more power than ever to study how genetic

instructions give us all life's diversity of shapes and forms.

CRISPR has made this kind of gene tweaking cheaper, faster, and more accurate than ever. |||||||gene modification|||||||

This really makes me wonder, if you have this ability to tweak how butterfly patterns end up coming out

can we get more control and actually design butterfly artwork of our own?

Make butterflies look the way we want to?

I think we will be able to, so yes, we can... but should we?

It's a new power, a new tool to harness nature, so we're responsible, we need to

do things that are relatively ethical, I would say.