Experimental therapy could repair mutations that causes genetic diseases



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November 13, 2018

November 13, 2018 by Mike Williams, Rice University

Rice University Bioengineers uses a magnetic field to activate nanoparticle-attached baculoviruses in a tissue. The viruses, which normally infect alfalfa looper moths, are modified to deliver gene-editing DNA code only to cells that are targeted with magnetic field-induced local transduction. Credit: Laboratory of Biomolecular Engineering and Nanomedicine / Rice University
A new technology that is related to moth-infecting viruses and nanomagnets could be used to reduce the risk of disease, which is known to cause sickle cell disease, muscular dystrophy and cystic fibrosis.

Rice University Bioengineer Gang Bao has combined magnetic nanoparticles with a viral container of a particular species of moth to deliver CRISPR / Cas9 payloads that modify genes in a specific tissue or organ with spatial control.
Because magnetic fields are simple to manipulate, unlike light, pass easily through tissue, and allow them to be used in the field of viral payloads by inactivated in blood.
The research appears in Nature Biomedical Engineering.
In nature, CRISPR / Cas9 bolsters microbes' immune systems by recording the DNA of invaders. That gives microbes the ability to recognize and attacking invaders, but scientists have tried to adapt to CRISPR / Cas9 to repair mutations that cause genetic diseases and to manipulate DNA in laboratory experiments.
CRISPR / Cas9 has the potential to halt hereditary disease – if scientists can get the genome-editing machines to the right cells inside the body. But roadblocks remain, especially in delivering the gene-editing payloads with high efficiency.
Bao said it would be necessary to edit cells in the body to treat many diseases. "But not enough," says Bao said. "Even if you inject the viral vector locally, it can leak to other tissues and organs, and that could be dangerous."

Gang Bao and his colleagues at Rice University are a combination of nanoparticles, magnets, and viruses that have a negative impact on their ability to deliver genetic therapy to people with inherited genetic diseases. Credit: Jeff Fitlow / Rice University
The delivery vehicle developed by Bao's group is based on a virus that infects Autographa californica, aka the alfalfa looper, a native moth to North America. The cylindrical baculovirus vector (BV), the payload-carrying part of the virus, is widely considered at up to 60 nanometers in diameter and 200-300 nanometers in length. That's big enough to transport more than 38,000 base pairs of DNA, which is enough to supply multiple gene-editing units to a target cell, Bao said.
He said the inspiration to combine BV and magnetic nanoparticles came from discussions with Rice postdoctoral researcher and co-lead author Haibao Zhu, who learned about the virus during a postdoctoral stint in Singapore but knew nothing about magnetic nanoparticles until he joined the Bao lab. The rice team has been experimenting with nanoparticles and anecdotal chemistry.

"We really did not know it, but we thought, 'worth a shot,'" Bao said.
The researchers use the magnetic nanoparticles to activate BV and deliver gene-editing payloads only where they are needed. To do this, they take advantage of an immune-system protein called C3 that normally inactivates baculoviruses.
"If we combine BV with magnetic nanoparticles, we can overcome this deactivation by applying the magnetic field," Bao said. "The beauty is that when we deliver it, gene editing occurs only at the tissue, or the part of the tissue, where we apply the magnetic field."
Application of the magnetic field allows BV transduction, the payload-delivery process that introduces gene-editing cargo into the target cell. The payload is also DNA, which encodes both a gene reporter and the CRISPR / Cas9 system.
In tests, the BV was loaded with green fluorescent proteins or firefly luciferase. Cells with the protein glowed brightly under a microscope, and experiments shown to be highly effective in the field of BV.
Other methods of treatment include CRISPR / Cas9 with adeno-associated viruses (AAV). "However, it is necessary to make BV transduction into the target cells more efficient," he said.

Explore further:
Are humans immune to the CRISPR-Cas9 gene editing scissors?

More information:
Haibao Zhu et al. Spatial control of in vivo CRISPR-Cas9 genome editing via nanomagnets, Nature Biomedical Engineering (2018). DOI: 10.1038 / s41551-018-0318-7

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