Scientists create albino lizards with CRISPR tech

Scientists create albino lizards with CRISPR tech”

So far, the CRISPR gene-editing system has been successfully used in bacteria, plants, mice, pigs, monkeys, and even humans - but never reptiles, due to their tricky reproductive methods.

A staff of scientists on the College of Georgia has efficiently carried out gene-editing on reptiles for the primary time, making a pair of albino lizards the scale of a human index finger.

"For quite some time we've been wrestling with how to modify reptile genomes and manipulate genes in reptiles, but we've been stuck in the mode of how gene editing is being done in the major model systems" mentioned Doug Menke, co-author of a paper that described the work in Cell Press on Tuesday.

As lizards have internal fertilization processes that can not be predicted, the traditional route of injecting CRISPR-Cas9 gene-editing reagents into freshly fertilized eggs or single-cell zygotes was not an option. In lizards, fertilization happens internally in the cloaca, but it's impossible to tell exactly when it takes hold. Also, the physiology of the fertilized eggs makes it hard for manipulating the embryos without damaging them.

But Menke and his research team noticed that the transparent membrane over the ovary allowed them to see all of the developing eggs, including which eggs were going to be ovulated and fertilized next. Instead, the researchers opted to inject CRISPR reagents directly into the unfertilized eggs within the ovaries.

"So we wanted to develop a new animal model to study fovea defects, and it turns out these lizards need high acuity vision, because when they hunt insects, they have fovea in their eyes". Scientists injected a solution targeting the tyrosinase gene into 146 oocytes inside 21 female brown anoles, a common lizard species in the Caribbean.

Reagents were injected into the eggs waiting to be fertilised and after a three-month wait for the baby lizards to hatch, the operation proved successful.

Not only did it work, but, to their surprise, the gene-edits ended up in both the maternal-line and paternal DNA, not just the former as they had predicted.

Researchers report producing the first gene-edited reptiles ever.

In some other model animals, CRISPR-Cas can have efficiencies up to 80% or higher, which would make the present 6% seem like a paltry amount, Menke explains.

That way any mutant lizards would be born albino, making it very obvious if the procedure worked. First, when the tyrosinase albinism gene is knocked out, it results in a loss of pigmentation without being lethal to the animal.

Lastly, people with albinism typically have issues with their imaginative and prescient, and researchers can use the index-finger-sized lizards as a mannequin to check how the gene impacts the event of the retina.

The anole lizards are ideally suited for this: their eyes have a fovea, a pit-like structure in the retina that underpins high-acuity vision, which humans share, but most of our main animal models lack. There is the potential of translating the gene-editing technique for use in other animals.

Scientists gene-edited anole lizards for a study published in the journal Cell Reports.

This work was supported by the National Science Foundation, a Society for Developmental Biology Emerging Models grant, an NIH training grant, and by an ARCS Foundation Scholarship.

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