New DNA repair method successfully repairs disease-causing gene mutation in patient's kidney cells

In a new study, researchers from the University of Bristol in the United Kingdom and the Caroline College in Sweden have successfully used a DNA repair tool that could lead to changes in the treatment of a debilitating inherited kidney disease in young peopl.

In the new study, the researchers describe how they constructed DNA repair tools to repair Podocin deficiency, a common genetic cause of inherited steroid-resistant nephrotic syndrome (SRNS.

Podocin is a protein on the surface of specific kidney cells that is essential for kidney functio.

These viruses are used as "Trojan horses" to enter cells that carry genetic mutation.

However, these viruses all share the same limitation, the limited space within the viral envelop.

This in turn limits their ability to deliver the "cargo" -- the DNA sequences needed for efficient gene repair -- which greatly limits their use in gene therap.

By applying synthetic biology techniques, a research team led by Dr Francesco Aulicino and Professor Imre Berger from the University of Bristol's School of Biochemistry has redesigned the baculovirus, an insect virus that is harmless to humans and no longer limited in "cargo" capacit.

"Baculovirus differs from LV, AV and AAV in that it lacks a hard shell to enclose the 'cargo' space," D.

This means that baculovirus could provide a more sophisticated toolbox for repairing genetic defects, making it more versatile than commonly used system.

First, the baculovirus must have the ability to enter human cells, which it usually does not hav.

"We decorate the baculovirus with proteins that allow it to enter human cells very efficiently," explains D.

The researchers then used their engineered baculovirus to deliver much larger pieces of DNA than before and integrated these into the genomes of a range of human cell.

The DNA in the human genome is made up of 3 billion base pairs, making up about 25,000 genes that encode proteins essential for cell functio.

The researchers used patient-derived podocytes that carry disease-causing errors in their genomes to demonstrate the applicability of their techniqu.

This reverses the pathogenic phenotype and restores Podocin on the podocyte surfac.

Rapid assembly of multifunctional DNA circuits for efficient baculovirus delivery in human cell.

"We have previously infected insect cells in vitro with baculovirus to produce recombinant proteins and study their structure and function," explains Pro.
Berge.

The method, called MultiBac, was developed in Berger's lab and is used worldwid.
In his lab, large multiprotein complexes with multiple subunits have been successfully fabricate.

By using our baculovirus delivery system and the latest editing technologies available, we can efficiently correct different sites on the genome simultaneousl.
Many mistake.

Co-author Professor Moin Saleem, from the University of Bristol, said: "SRNS is one of the more common genetic disorders affecting the kidney.
SRNS ischaracterized by early onset kidney failure that severely reduces the quality of life of patient.
These results are very encouragin.
The new approach pioneered by Berger's team holds promise for treating not only SRNS but also a range of other genetic diseases of the kidne.
It is also hoped that in these diseases, efficient gene repair is not feasible with current technolog.
The clinical application of the vector system is still a long way off, but we believe the advantages it offers make it a very valuable research endeavo.
"