Researchers master gene editing technique in mosquito that transmits deadly diseases

March 26, 2015
Researchers master gene editing technique in mosquito that transmits deadly diseases
Mosquito larvae from two different lines fluoresce in different colors thanks to genetic tags that were inserted using the CRISPR-Cas9 gene editing system. Credit: Vosshall Laboratory

Traditionally, to understand how a gene functions, a scientist would breed an organism that lacks that gene - "knocking it out" - then ask how the organism has changed. Are its senses affected? Its behavior? Can it even survive? Thanks to the recent advance of gene editing technology, this gold standard genetic experiment has become much more accessible in a wide variety of organisms. Now, researchers at Rockefeller University have harnessed a technique known as CRISPR-Cas9 editing in an important and understudied species: the mosquito, Aedes aegypti, which infects hundreds of millions of people annually with the deadly diseases chikungunya, yellow fever, and dengue fever.

Researchers led by postdoctoral fellow Benjamin J. Matthews adapted the CRISPR-Cas9 system to Ae. aegypti and were able to efficiently generate targeted mutations and insertions in a number of genes. The immediate goal of this project, says Matthews, is to learn more about how different genes help the species operate so efficiently as a disease vector, and create new ways to control it. "To understand how the actually transmits disease," says Matthews, "you have to learn how she finds humans to bite, and how she chooses a source of water to lay her eggs. Once you have that information, techniques for intervention will come."

In the study, published March 26 in Cell Reports, Matthews and research assistant Kathryn E. Kistler, both in Leslie B. Vosshall's Laboratory of Neurogenetics and Behavior, adapted the CRISPR-Cas9 system to introduce precise mutations in Ae. aegypti. Previously, to create these types of mutations, scientists relied on techniques that used engineered proteins to bind to specific segments of DNA they wanted to remove, a process that was both expensive and unreliable. CRISPR-Cas9, in contrast, consists of short stretches of RNA that bind to specific regions of the genome where a protein, Cas9, cleaves the DNA. Scientists have been studying how RNA binds to DNA for decades and "the targeting is done with rules that we have a good handle on," says Matthews, which makes it easy to reprogram CRISPR-Cas9 to target any gene.

"This amazing technique has worked in nearly every organism that's been tried," says Vosshall, who is Robin Chemers Neustein Professor and a Howard Hughes Medical Institute investigator. "There are lots of interesting animal species out there that could not be studied using genetics prior to CRISPR-Cas9, and as a result this technique is already revolutionizing biology."

This work opens the door to learning more about the role of specific genes the Vosshall lab suspects may help mosquitoes propagate, perhaps by finding the perfect spot to lay their eggs. Their protocols will likely also help other scientists apply the same technique to study additional organisms, such as agricultural pests or mosquitoes that carry malaria.

"Before starting this project, we thought it would be difficult to modify many genes in the mosquito genome in a lab setting" Matthews says. "With a little tweaking, we were able to make this technique routine and it's only going to get easier, faster, and cheaper from here on out."

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JVK
1 / 5 (3) Mar 26, 2015
"This work opens the door to learning more about the role of specific genes the Vosshall lab suspects may help mosquitoes propagate..."

My comment: Other works from her group close the door on theories about mutations, natural selection, and the evolution of biodiversity by differentiating between mutations and nutrient-dependent RNA-directed DNA methylation linked to RNA-mediated amino acid substitutions that differentiate the cell type of all individuals of all species from microbes to man.

orco mutant mosquitoes lose strong preference for humans and are not repelled by volatile DEET http://www.ncbi.n...3696029/

Evolution of mosquito preference for humans linked to an odorant receptor
http://www.nature...964.html

See also: Use of a structural alphabet to find compatible folds for amino acid sequences http://dx.doi.org...pro.2581

http://www.ncbi.n...24693353
ayesdi_fdesay
not rated yet Mar 26, 2015
Amazing stuff. Are we able to edit the genes of live organisms with CRISPR-Cas9 at this point in time?
JVK
1 / 5 (3) Mar 26, 2015
Live organisms do it themselves via RNA-mediated events during their life history transitions

Clinically Actionable Genotypes Among 10,000 Patients With Preemptive Pharmacogenomic Testing http://www.medsca...24253661

Oppositional COMT Val158Met effects on resting state functional connectivity in adolescents and adults http://dx.doi.org...4-0895-5

From Fertilization to Adult Sexual Behavior http://www.hawaii...ion.html

Our model was linked to insects in: Organizational and activational effects of hormones on insect behavior http://www.ncbi.n...10980296

It was linked to their life history transitions in Honey bees as a model for understanding mechanisms of life history transitions http://www.ncbi.n...15925525

What we detailed in the context of our section on molecular epigenetics does not change across species.
Vietvet
3.7 / 5 (3) Mar 27, 2015
Amazing stuff. Are we able to edit the genes of live organisms with CRISPR-Cas9 at this point in time?


@ayesdi fdeasy
I can't answer your question but beware anything JVK writes. He is a self serving wanta be pseudo scientist. He is well known here as a self proclaimed creationist and college drop-out.
He sells a "perfume" based on totally debunked pseudo sconce.

Over and over he misrepresents the work of real scientists.

He thinks linking to creationist BLOGS bolsters his delusional beliefs without realizing that readers here recognize him for the fool he is.

The very first thread I read here with JVK's comments I pictured him gazing admirably in a mirror while wearing a skirt , it was the only explanation for his narcissism and shrillness.
anonymous_9001
5 / 5 (3) Mar 27, 2015
Life history transitions are due to alternative splicings and altered gene transcription levels.

However, gene editing is entirely different. Genes are not changed through splicing or changing the amount of RNA derived from them.
Lex Talonis
not rated yet Mar 27, 2015
With this tech, you could really fuck with mosquitos...

Like give them legs / wings / harpoons etc., twice as long, bodies twice as heavy and half the brain they currently have..

Graft in the flies avoidance of the dark, add in 8 sets of eyes like a spider, glow worm genes, the ability to spontaneously combust when having sex...

And addiction to alcohol etc..

You could be sooooo cruel to generations of them in thousands of ways...
JVK
1 / 5 (3) Mar 27, 2015
JVK
What we detailed in the context of our section on molecular epigenetics does not change across species.,


anonymous_9001 (aka Andrew Jones)
Life history transitions are due to alternative splicings and altered gene transcription levels.


I already claimed that what we detailed in the context of our section on molecular epigenetics was extended to:
1) insects in 2000 and
2) to the life history transitions of the honeybee model organism in 2005 and
3) to what is known about the biophysically constrained RNA-mediated chemistry of protein folding in species from microbes to man in 2014.

See this 2 minute video: https://www.youtu...G_9EEeeA
See this 5.5 minute video: http://youtu.be/DbH_Rj9U524

Ask the anonymous fool (aka Andrew Jones) what he is claiming when he says:
...gene editing is entirely different. Genes are not changed through splicing or changing the amount of RNA derived from them.
JVK
1 / 5 (3) Mar 27, 2015
Over and over he misrepresents the work of real scientists.


http://rna-mediated.com/ "Here you will find information that links physics, chemistry, and molecular epigenetics via RNA-mediated events such as the de novo creation of olfactory receptor genes..."

See also: http://perfumingthemind.com/ "James V. Kohl was the first to accurately conceptualize human pheromones, and began presenting his findings to the scientific community in 1992. He continues to present to, and publish for, diverse scientific and lay audiences, while constantly monitoring the scientific presses for new information that is relevant to the development of his initial and ongoing conceptualization of human pheromones."

Vosshall's works and detailed explanations of biologically-based cause and effect link what is currently known about nutrient-dependent RNA-mediated amino acid substitutions to the pheromone-controlled physiology of reproduction in species from microbes to man.
JVK
1 / 5 (3) Mar 27, 2015
Life history transitions are due to alternative splicings and altered gene transcription levels.


See the molecular epigenetics section of our 1996 H&B review:
http://www.hawaii...ion.html
"Small intranuclear proteins also participate in generating alternative splicing techniques of pre-mRNA and, by this mechanism..."

Our model has been extended across species by many others.
http://genomebiol...15/1/401]http://genomebiol...15/1/401[/url]
"...the interactions between pre-mRNA and proteins fine-tune alternative splicing in a manner that can gradually create new protein functionalities without the need to create additional genes and without affecting existing proteins [4-6]." http://genomebiol...15/1/401]http://genomebiol...15/1/401[/url]

See also: Nutrient-dependent/pheromone-controlled adaptive evolution: a model. http://www.ncbi.n...24693353 and a fool's criticisms of the model http://www.ncbi.n...24959329
JVK
1 / 5 (3) Mar 27, 2015
Editorial for "Regulatory RNAs in the nervous system"

"Particularly well-characterized is the role of microRNAs (miRNAs) in the post-transcriptional regulation of gene expression. MicroRNAs are short(~21 nt) nc-RNAs that arise from processing of a long primary transcript via a complex and well-described biosynthetic process. MicroRNAs bind to mRNAs (usually in the 3′untranslated region) and regulate gene expression by repressing mRNA translation and/or inducing degradation of the target mRNA. Up to now, several thousands of miRNAs have been predicted and identified in animals, plants and viruses (www.mirbase.org) and some microRNAs are highly conserved, facilitating the analysis of microRNA in non-model species."

http://journal.fr...w13-2015
JVK
1 / 5 (3) Mar 27, 2015
Regulatory RNAs in the Nervous System http://journal.fr...pic/1259
34 articles; 160 authors

My invited review of nutritional epigenetics: Nutrient-dependent pheromone-controlled ecological adaptations: from atoms to ecosystems
http://figshare.c...s/994281

Excerpt: "More substantial support for epigenetic effects on cell type differentiation comes from what has been learned during the past decade about the role of small non-coding RNA molecules. The small non-coding RNA molecules are called microRNAs (miRNAs). MiRNAs alter intercellular [correction: intracellular] signaling by changing the balance between miRNAs and messenger RNA (mRNA) . The changes are linked to health and to pathology (Mori et al., 2014)."

Clarification: Nutrient-dependent RNA-mediated amino acid substitutions are linked to health; mutations are linked to physiopathology.
JVK
1 / 5 (3) Mar 27, 2015
See also: Mitochondrial genetic disorders http://www.mlo-on...ders.php

The links to nutrient-dependent RNA-mediated cell type differentiation are not very clear in this article, except in the context of personalized medicine, which links nutrient-dependent metabolic networks and genetic networks in all genera via the biophysically constrained chemistry of RNA-directed DNA methylation and RNA-mediated amino acid substitutions.

However, other excellent articles by this author make the facts perfectly clear. John Brunstein has detailed many aspects of how thermodynamic cycles of protein biosynthesis and degradation link the nutrient-dependent microRNA/messenger RNA balance to health and physiopathology via conserved molecular mechanisms (e.g., from atoms to ecosystems).

He provides detailed representations of biologically-based cause and effect with unparallelled clarity.
anonymous_9001
5 / 5 (3) Mar 27, 2015
By "amino acid substitution" are you referring to substitutions that occur during RNA editing after transcription?
JVK
1 / 5 (3) Mar 27, 2015
Excerpt: "This work opens the door to learning more about the role of specific genes the Vosshall lab suspects may help mosquitoes propagate..."

Contact Leslie Vosshall, remind her that she claimed my work was pseudoscience. Ask her any questions you have about "amino acid substitutions" and when or how cell type differentiation occurs.

As you may recall, I have a detailed model that placed amino acid substitutions into the context of biologically-based cause and effect based on what we reported about RNA-mediated cell type differentiation in the molecular epigenetics section of our 1996 Hormones and Behavior review. From Fertilization to Adult Sexual Behavior http://www.hawaii...ion.html

I mentioned the model (above):
Nutrient-dependent/pheromone-controlled adaptive evolution: a model.
http://www.ncbi.n...24693353

and your criticisms of it http://www.ncbi.n...24959329
JVK
1 / 5 (3) Mar 27, 2015
Relatedness, Conflict, and the Evolution of Eusociality
http://journals.p....1002098
Conclusion: "To have multiple theoretical approaches converging on similar results attests to the robustness of social evolution theory."

The criticisms of my model by Andrew Jones (aka anonymous_9001) attest to the robustness of pseudoscientific nonsense taught as if the ridiculous theories based on the definition of mutation and assumptions about how long it took for one species to become a different species could be compared to the honeybee model [or any other insect model] of nutrient-dependent pheromone-controlled RNA-mediated biologically-based cell type differentiation.

http://www.ncbi.n...24693353 Conclusion: "Minimally, this model can be compared to any other factual representations of epigenesis and epistasis for determination of the best scientific 'fit'."

(e.g. the robustness of social evolution theory?)
anonymous_9001
5 / 5 (3) Mar 27, 2015
The honeybee model concerns how different castes can be derived from identical genomes, not how genomes change over time.
JVK
1 / 5 (3) Mar 27, 2015
In your model, what causes the epigenesis that leads to epistasis or anything else that you think causes genomes to change over time?

I reiterate: "As you may recall, I have a detailed model that placed amino acid substitutions into the context of biologically-based cause and effect based on what we reported about RNA-mediated cell type differentiation in the molecular epigenetics section of our 1996 Hormones and Behavior review."

When are you going to explain HOW species evolve?

http://rna-mediat...animals/
anonymous_9001
5 / 5 (3) Mar 28, 2015
Epigenesis is the developmental process an organism goes through as it matures. Epistasis is an interaction of two or more genes to determine a single phenotype.

Development over the lifetime of a single organism is not the same as a population changing over subsequent generations. I'm not sure why you're trying to equate the two. They're completely separate processes.
JVK
1 / 5 (2) Mar 28, 2015
When are you going to explain HOW species evolve?

http://phys.org/n...firstCmt
JVK
1 / 5 (2) Mar 28, 2015
Here are publications by others who are "Combating Evolution to Fight Disease"
http://www.scienc...88.short

Note: They are publishing works that refute neo-Darwinian theory at the same time as Andrew Jones refuses to tell us HOW species evolve.
http://www.labome...583.html

Maternal and fetal genetic associations of PTGER3 and PON1 with preterm birth
Kelli K Ryckman

Epistatic interactions in genetic regulation of t-PA and PAI-1 levels in a Ghanaian population
Nadia M Penrod

A population-based study in Ghana to investigate inter-individual variation in plasma t-PA and PAI-1 Scott M Williams

Geographic ancestry and markers of preterm birth Scott M Williams

The use of animal models in the study of complex disease: all else is never equal or why do so many human studies fail to replicate animal findings? Scott M Williams
JVK
1 / 5 (2) Mar 28, 2015
JVK: "what causes the epigenesis that leads to epistasis"

I'm not sure why you're trying to equate the two.


Obviously, I am not trying to equate the two. I asked you about a cause and effect relationship. It is required to link the epigenetic landscape to the physical landscape of DNA in the organized genomes of species from microbes to man via the biophysically constrained chemistry of protein folding and the conserved molecular mechanisms of RNA-mediated amino acid substitutions that I have detailed with examples in my model.

Nutrient-dependent/pheromone-controlled adaptive evolution: a model.
http://www.ncbi.n...24693353

Development over the lifetime of a single organism is not the same as a population changing over subsequent generations.


What causes the change(s) during the lifetime of a single organism that cause population change(s) over subsequent generations?

See also" http://www.ncbi.n...24693349

anonymous_9001
5 / 5 (2) Mar 28, 2015
They are publishing works that refute neo-Darwinian theory


I've already pointed out to you how that paper describes mutagenesis. They don't refute neo-Darwinism in any way, shape, or form.

Obviously, I am not trying to equate the two


You based your evolutionary model on the the honeybee model. How is that not equating the two?

What causes the change(s) during the lifetime of a single organism that cause population change(s) over subsequent generations?


You are so far off the mark here... Based on that question, you don't understand the honeybee model in the first place. It describes how different castes can be derived from the SAME genetics.

A bee larva has the ability to develop into all the different castes (worker, drone, queen). No changes to the genome are made during this development process. That's the entire point of epigenesis. However, between subsequent generations, genomes do change.
JVK
1 / 5 (1) Mar 29, 2015
You want to separate mutations from natural selection; separate selection from evolution; and now you want to separate epigenesis from epistasis.

I've linked physics to the chemistry of protein folding and the conserved molecular mechanisms of cell type differentiation exemplified in the honeybee model organism of nutrient-dependent pheromone-controlled RNA-directed DNA methylation and RNA-mediated amino acid substitutions.

Which one of your beloved family members are you willing to offer as the first sacrifice to your ignorance of cell type differentiation?

http://www.ncbi.n...3960065/
"The honeybee already serves as a model organism for studying human immunity, disease resistance, allergic reaction, circadian rhythms, antibiotic resistance, the development of the brain and behavior, mental health, longevity, diseases of the X chromosome, learning and memory, as well as conditioned responses to sensory stimuli (Kohl, 2012)."

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