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.”
Could this protein protect people against coronary artery disease?
17.11.2017 | University of North Carolina Health Care
Microbial resident enables beetles to feed on a leafy diet
17.11.2017 | Max-Planck-Institut für chemische Ökologie
The formation of stars in distant galaxies is still largely unexplored. For the first time, astron-omers at the University of Geneva have now been able to closely observe a star system six billion light-years away. In doing so, they are confirming earlier simulations made by the University of Zurich. One special effect is made possible by the multiple reflections of images that run through the cosmos like a snake.
Today, astronomers have a pretty accurate idea of how stars were formed in the recent cosmic past. But do these laws also apply to older galaxies? For around a...
Just because someone is smart and well-motivated doesn't mean he or she can learn the visual skills needed to excel at tasks like matching fingerprints, interpreting medical X-rays, keeping track of aircraft on radar displays or forensic face matching.
That is the implication of a new study which shows for the first time that there is a broad range of differences in people's visual ability and that these...
Computer Tomography (CT) is a standard procedure in hospitals, but so far, the technology has not been suitable for imaging extremely small objects. In PNAS, a team from the Technical University of Munich (TUM) describes a Nano-CT device that creates three-dimensional x-ray images at resolutions up to 100 nanometers. The first test application: Together with colleagues from the University of Kassel and Helmholtz-Zentrum Geesthacht the researchers analyzed the locomotory system of a velvet worm.
During a CT analysis, the object under investigation is x-rayed and a detector measures the respective amount of radiation absorbed from various angles....
The quantum world is fragile; error correction codes are needed to protect the information stored in a quantum object from the deteriorating effects of noise. Quantum physicists in Innsbruck have developed a protocol to pass quantum information between differently encoded building blocks of a future quantum computer, such as processors and memories. Scientists may use this protocol in the future to build a data bus for quantum computers. The researchers have published their work in the journal Nature Communications.
Future quantum computers will be able to solve problems where conventional computers fail today. We are still far away from any large-scale implementation,...
Pillared graphene would transfer heat better if the theoretical material had a few asymmetric junctions that caused wrinkles, according to Rice University...
15.11.2017 | Event News
15.11.2017 | Event News
30.10.2017 | Event News
17.11.2017 | Physics and Astronomy
17.11.2017 | Health and Medicine
17.11.2017 | Studies and Analyses