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PRINCE: A Small-Molecule Switch for Safer Gene Editing
PRINCE: A Small-Molecule Switch for Safer Gene Editing
Although gene editing has enormous clinical promise, it still faces many obstacles that must be overcome before broad translation. For example, the genome editing field is continuously working to increase the safety of the technique. One way to do that is to control the duration of gene editing activity. However, achieving precise temporal control over these platforms is challenging.
Scientists currently lack tools that can precisely control the duration of gene editing therapies, to halt their effects after a few months or years. Researchers have developed several potential solutions, including degradable protein- or RNA-based systems, but existing approaches face limitations such as only being applicable in the liver.
Now, a new gene editing system, named PRINCE, with inducible nuclease proteins and guide RNAs, enables researchers to control the duration and specificity of gene therapies precisely with small molecules—potentially addressing a longstanding safety concern in the field.
This work is published in Science Translational Medicine in the paper, “Coordinated regulation using small-molecule drugs enables controlled therapeutic genome editing and enhanced genomic precision in situ.“
In the PRINCE CRISPR-Cas gene editing system, expression of the nuclease and the guide RNA is separately inducible by two approved small molecule drugs, enabling both temporal control and reduced off-target editing.
The platform was stable for as long as two years after genomic integration in cultured human cells. In addition, a smaller system, called Little Prince, showed promising signs of efficacy in humanized mouse models of elevated cholesterol levels and age-related macular degeneration. Little Prince uses compact nucleases that can be delivered in a single adeno-associated viral vector (AAV) for delivery.
In two mouse models, Little Prince reduced excessive cholesterol levels in mice with genetic hypercholesterolemia and showed signs of reducing lesion size in rodents with laser-induced choroidal neovascularization—a model of age-related macular degeneration.
More specifically, Little Prince “ameliorated pathological phenotypes of hypercholesterolemia (average reductions of 45% and 47% in serum total cholesterol and low-density lipoprotein cholesterol, respectively) and neovascular age-related macular degeneration, with significantly reduced lesion size and leakage (P < 0.0001).”
The system produced fewer off-target edits and lower off-target editing frequencies than constitutive nuclease expression, highlighting the utility of precise temporal control.
“PRINCE and Little Prince also provide […] capabilities that might also be useful for research objectives that include lineage tracing and conditional genetic engineering work,” the authors noted. These results, they asserted, position PRINCE and Little Prince as controlled genome editing platforms with potential for in vivo, particularly in situ, therapeutic applications.
The post PRINCE: A Small-Molecule Switch for Safer Gene Editing appeared first on GEN – Genetic Engineering and Biotechnology News.
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New webinar: Tackling drug discovery challenges in cancer research

Hosted by Drug Discovery World and supported by Sartorius and BioIVT, this webinar will explore the opportunities and challenges that exist within cancer research drug discovery and development.
You will hear from Dr Sudha Rao, Chief Scientific Officer of Kazia Therapeutics, Karol Budzik, PhD, Business Development Associate at Vyriad Therapeutics and Lars van der Veen, Chief Scientific Officer at iOnctura.
Presentations will cover how cancer treatments have shifted towards reprogramming the biology driving tumour growth, immune escape and treatment resistance, the trajectory that in vivo CAR-T treatments are taking, and how challenging tumours burdened by stroma and immune-mediated resistance can be tackled.
Q&A with the speakers follows the presentations.
The post New webinar: Tackling drug discovery challenges in cancer research appeared first on Drug Discovery World (DDW).
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Psilocybin proves promising in neuropathic pain mouse study
Amid the rise of psychedelics in the mental health space, researchers have begun to explore psilocybin as a treatment for chemotherapy-induced peripheral neuropathy.
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New Spectrometry Technique Could Aid Formulation Development
New Spectrometry Technique Could Aid Formulation Development
A new technique combining two forms of spectrometry could help biopharmaceutical companies improve their choice of formulation buffer for antibody manufacturing by revealing how molecular forms and three-dimensional shapes of complex biologics respond to their environment. That’s the view of Christian Bleiholder, PhD, a professor at Florida State University who helped develop the technique.
According to Bleiholder, what happens structurally when a complex biological molecule, such as an antibody or viral spike protein, binds to its target is currently poorly understood.
“This is where [this approach] can help with the bioprocessing and formulation,” he says, as structural changes “can affect the lifespan [of the product] and lead to issues, such as aggregation.”
Because antibodies are complex, existing techniques tend to be powerful at different levels of complexity, he explains. Mass spectrometry is particularly powerful for distinguishing molecular composition, while structural approaches such as X-ray crystallography and cryo-electron microscopy can provide high-resolution structural information.
The challenge is understanding the link between these things within a heterogeneous sample, he says.
To overcome this, Bleiholder and his team worked with Bruker Daltonics to develop Tandem-Trapped Ion Mobility Spectrometry (Tandem-TIMS). This combines tandem ion mobility spectrometry with tandem mass spectrometry to disentangle three overlapping layers of molecular complexity: molecular form, three-dimensional shape, and binding or assembly state, he says.
He explains that, if the proteins have different structures, they can be characterized with tandem ion mobility spectrometry, and then mass spectrometry can be used to look at their molecular forms and binding states.
Going forward, Bleiholder hopes the technique can be used for formulation development but also earlier, during drug discovery of new products, such as multi-specific antibodies, to determine which molecular states are important and how those change when a biologic engages its target. He also plans to look at automating the technique.
Bleiholder spoke about using Tandem-TIMS at the Bioprocessing Summit in Boston earlier this year.
The post New Spectrometry Technique Could Aid Formulation Development appeared first on GEN – Genetic Engineering and Biotechnology News.
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