An optimized technique allows B cells to be transfected with extraneous DNA without the use of viruses
The introduction of foreign DNA into human cells through a process known as ‘transfection’ allows scientists to study gene expression in the laboratory and enables clinicians to treat genetic diseases. The methods commonly used for this procedure work for most cell types, except when it comes to B cells — a group of infection-fighting white blood cells in the immune system that have proven extremely difficult to transfect without the use of viruses. Viruses, however, pose a number of safety issues.
A team led by scientists at the A*STAR Bioprocessing Technology Institute and the A*STAR Institute of High Performance Computing has now developed a non-viral strategy to deliver DNA into this intractable cell type. By optimizing a technique termed sonoporation, the researchers managed to introduce genes into B cells with high rates of success1.
“Our work is the first to demonstrate the use of sonoporation as an alternative, non-viral method for stable and highly efficient transfection of recalcitrant B cell lines,” says biomedical engineer and study leader Andre Boon-Hwa Choo.
Sonoporation combines ultrasonic sound frequencies and tiny gas-filled bubbles to generate transient pores in the cell membrane through which DNA can travel. Choo and his colleagues tweaked the acoustic energy levels and microbubble concentrations to deliver a circular piece of DNA that they could track visually in a trio of human B cell lines.
In one cell line, for example, the researchers achieved around 43 per cent transfection efficiency through sonoporation, compared to just 3 per cent with a conventional transfection method called lipofection (see image). Through further selection techniques, the researchers enriched the population of transfected B cells to more than 70 per cent. They achieved similarly impressive results with the two other B cell lines.
According to Charlene Li Ling Yong, co-first-author of the study along with Dave Siak-Wei Ow, the sonoporation-based transfection technique can now be used in the laboratory to better understand how B cells regulate immune responses against pathogens. “It allows scientists to elucidate the biological pathways of immune responses,” says Yong.
Numerous clinical research teams are also pursuing B-cell-based gene therapies to induce tolerance against autoimmune diseases. The method described in the current study could come in particularly handy for treatments in the human body — without any of the adverse effects of viral-mediated gene therapy. “Sonoporation has the potential to be applied in vivo,” Ow says. “It offers a safer and noninvasive alternative to existing gene therapies.”
Scientists uncover the role of a protein in production & survival of myelin-forming cells
19.07.2018 | Advanced Science Research Center, GC/CUNY
NYSCF researchers develop novel bioengineering technique for personalized bone grafts
18.07.2018 | New York Stem Cell Foundation
A new manufacturing technique uses a process similar to newspaper printing to form smoother and more flexible metals for making ultrafast electronic devices.
The low-cost process, developed by Purdue University researchers, combines tools already used in industry for manufacturing metals on a large scale, but uses...
For the first time ever, scientists have determined the cosmic origin of highest-energy neutrinos. A research group led by IceCube scientist Elisa Resconi, spokesperson of the Collaborative Research Center SFB1258 at the Technical University of Munich (TUM), provides an important piece of evidence that the particles detected by the IceCube neutrino telescope at the South Pole originate from a galaxy four billion light-years away from Earth.
To rule out other origins with certainty, the team led by neutrino physicist Elisa Resconi from the Technical University of Munich and multi-wavelength...
For the first time a team of researchers have discovered two different phases of magnetic skyrmions in a single material. Physicists of the Technical Universities of Munich and Dresden and the University of Cologne can now better study and understand the properties of these magnetic structures, which are important for both basic research and applications.
Whirlpools are an everyday experience in a bath tub: When the water is drained a circular vortex is formed. Typically, such whirls are rather stable. Similar...
Physicists working with Roland Wester at the University of Innsbruck have investigated if and how chemical reactions can be influenced by targeted vibrational excitation of the reactants. They were able to demonstrate that excitation with a laser beam does not affect the efficiency of a chemical exchange reaction and that the excited molecular group acts only as a spectator in the reaction.
A frequently used reaction in organic chemistry is nucleophilic substitution. It plays, for example, an important role in in the synthesis of new chemical...
Optical spectroscopy allows investigating the energy structure and dynamic properties of complex quantum systems. Researchers from the University of Würzburg present two new approaches of coherent two-dimensional spectroscopy.
"Put an excitation into the system and observe how it evolves." According to physicist Professor Tobias Brixner, this is the credo of optical spectroscopy....
13.07.2018 | Event News
12.07.2018 | Event News
03.07.2018 | Event News
20.07.2018 | Power and Electrical Engineering
20.07.2018 | Information Technology
20.07.2018 | Materials Sciences