Physical Gene Transfer: Advances in Genetic Research
The development of physical methods for gene transfer dates back to the early 1980s, with direct microinjection emerging as one of the first techniques used to introduce nucleic acids into cells. Microinjection involves the precise delivery of DNA or RNA into the cytoplasm or nucleus of cultured cells using a fine needle. Despite its labor-intensive nature, microinjection has been widely used for introducing foreign DNA into Xenopus oocytes and Drosophila embryos. Although it exhibits relatively low efficiency compared to other transfection approaches, microinjection remains a valuable tool for transferring genetic material into embryonic stem cells for the production of transgenic organisms, particularly in agricultural biotechnology and genetic studies involving farm animals. More recently, microinjection has gained renewed interest in gene-editing applications, particularly in delivering CRISPR-Cas9 components into zygote pronuclei to generate genetically modified animal models such as knockout pigs.
Electroporation is another widely utilized physical method for gene transfer. This technique applies an electrical pulse to temporarily disrupt the cell membrane, creating transient pores that facilitate the uptake of nucleic acids. While electroporation is highly effective, its success depends on careful optimization of pulse duration and intensity to balance transfection efficiency with cell viability. Due to the substantial cell death associated with this method, it often requires a larger number of cells than chemical transfection approaches. However, modern electroporation systems now allow for targeted nucleic acid delivery to the nucleus, significantly improving the efficiency of gene transfer into primary and stem cells. Recent advancements have integrated electroporation with microfluidic devices to facilitate high-throughput gene delivery by physically constricting cells prior to electrical pulsing, further enhancing transfection outcomes.
Another physical method used for gene transfer is biolistic particle delivery, or particle bombardment, which utilizes high-velocity microprojectiles to transfer nucleic acids into recipient cells by penetrating the cell membrane. This method has been successfully employed for gene transfer in both cultured cells and in vivo applications. Initially, the high cost of particle bombardment instrumentation limited its widespread adoption, with primary applications in agricultural genetics. However, the emergence of nanoparticle-based gene delivery has significantly expanded its use. Nanoparticles, defined as synthetic materials with at least one dimension under 100 nanometers, exhibit high membrane permeability, allowing for efficient nucleic acid delivery through covalent or non-covalent interactions with DNA and RNA. These properties have facilitated their application as an alternative to traditional pronuclear microinjection and viral vectors in transgenic animal research. A variation of this technique, known as magnetofection, utilizes iron oxide nanoparticles to enhance gene transfer efficiency in the presence of strong magnetic fields, representing an innovative approach to improving nucleic acid delivery.
The continued evolution of physical gene transfer methods, particularly in conjunction with emerging nanotechnology and microfluidics, has significantly enhanced the precision and efficiency of genetic manipulation. These methods provide valuable alternatives to chemical and viral-based transfection techniques, offering new avenues for functional genomics, gene therapy, and the development of transgenic models across various research domains.
Altogen Laboratory Research Services, Biology CRO Services
We provide a streamlined service for the generation of stably expressing cell lines within 28 days, enabling long-term expression of a vector or gene of interest in your chosen cell model. This process involves the transformation of either client-provided cell lines or one of our 120 in-house established cell lines and primary cells, ensuring broad applicability across diverse research fields. Stable expression is achieved through the integration or episomal retention of plasmid DNA constructs, which must include at least one antibiotic resistance gene to facilitate selective pressure during drug-based selection. Our optimized transfection protocols, coupled with rigorous selection and clonal expansion strategies, ensure high-efficiency integration, minimal off-target effects, and robust, reproducible gene expression. These stably modified cell lines serve as powerful tools for functional genomics, protein production, drug screening, and therapeutic development… Continue Reading
An Advanced Non-Viral Transfection Reagent for Central Nervous System Gene Delivery
The efficient delivery of nucleic acids into the central nervous system (CNS) of small animal models remains a significant challenge due to the limitations of conventional gene delivery methods. Standard non-viral gene carriers often exhibit insufficient transfection efficiency, while viral-based approaches, despite their effectiveness, are associated with considerable drawbacks, including complex production processes, safety concerns, and the need for stringent biosafety measures.
Altogen Biosystems in vivo transfection reagent represents a novel, non-viral transfection reagent specifically designed to overcome these challenges by enabling safe, efficient, and reproducible nucleic acid delivery into the CNS. Its unique formulation allows targeted transfection of neural cells within specific brain regions following stereotaxic injection, offering a highly effective approach for gene modulation studies. Unlike viral-based methods, Altogen Biosystems in vivo transfection reagent exhibits low immunogenicity, ensuring minimal immune response and reduced cytotoxicity. Furthermore, it supports both rapid and sustained transgene expression, making it a valuable tool for neurobiological research, gene therapy investigations, and functional studies of neural circuits.
Quality Control and Performance Standards
Each lot of Altogen Biosystems in vivo transfection reagent undergoes stringent quality control, ensuring sterility through thioglycolate assays with no bacterial or fungal contamination for 14 days. Biological activity is validated via in vitro transfection assays, maintaining at least 80% of the efficiency of a reference lot. Additionally, endotoxin levels are strictly controlled, remaining below 0.1 EU/mL to meet in vivo-grade safety standards.
Rat brain–computer interface. Delivery system with BMI
Altogen Biosystems has developed a rat brain–computer interface (BCI) using its proprietary Altogen® delivery system, integrating high-bandwidth brain chips to establish real-time communication between neural circuits and computational systems. This technology enables precise neuromodulation and high-resolution neural data acquisition, facilitating research in neurodegenerative diseases, cognitive function, and neuroprosthetic applications. Successfully tested in rat models of Alzheimer’s disease, the system allows for studying disease progression and therapeutic interventions. Future advancements aim to enhance signal processing, expand applications to other neurological disorders, and optimize biocompatibility for long-term integration… Continue Reading
The species we use:
One of the mammalian species is the laboratory rat, which has been researched the most extensively. Its usage has been reported in more than one million papers, and it has been used in a wide variety of medically significant fields. Its popularity may be attributed to its size, fecundity, behavior, and the simplicity with which surgical operations, tissue sampling, and routine laboratory management can be performed. On the other hand, due to the fact that technology to make genetic models of rats by targeting genes has not been successfully developed, the mouse has emerged as the most extensively used animal model.
Benefits of Rat Models Over Mouse Models
Rat models offer several significant advantages over mouse models for xenograft studies. Due to their larger body size, surgical manipulation and imaging procedures are easier to perform, allowing for more precise interventions and improved visualization. In terms of cardiovascular physiology and drug metabolism, rats exhibit greater similarity to humans than mice, making them a more relevant model for pharmacological and toxicological research. Additionally, when xenografts are implanted into specific tissues such as the prostate or brain, tumors in rats tend to grow larger, providing more tissue for analysis and enhancing the accuracy of experimental outcomes. Rats are also preferred for studies on memory and cognition, as their behavioral complexity and learning capabilities offer a superior model for neurological research. Furthermore, in a single experimental model, rats enable the simultaneous evaluation of both drug efficacy and toxicology, making them a valuable tool for preclinical research and translational medicine.
The importance of mice in research
Mice are important in research because they are small, easily manipulated, and have a genetic makeup similar to humans. Additionally, they are relatively cheap to maintain and breed.
Mice have been used in research for centuries and have been instrumental in furthering our understanding of biology and disease. For example, they were used to develop the smallpox vaccine and to study the effects of radiation.
Today, mice are still used in a wide variety of research projects, ranging from studying cancer to understanding the basic workings of the brain. They are also being used to develop new treatments for a variety of diseases, such as Alzheimer’s and Parkinson’s.
Overall, mice are an essential tool in the scientific world and will continue to play a vital role in research for years to come.
The advantages of using mice in research
Mice are one of the most commonly used animals in scientific research. There are several reasons for this, including their small size, their short life span, and the fact that they are easy to care for and breed in captivity. Additionally, mice are physiologically similar to humans in many ways, making them a good model for studying human diseases.
Mice are particularly well suited for genetic research. Their small size makes it possible to house large numbers of them in a small space, and their short life span means that researchers can observe the effects of genetic changes over several generations in a relatively short period of time. Additionally, because mice can be bred to have specific genetic characteristics, they are often used to study the effects of particular genes on health and disease.
Overall, mice are a valuable tool for scientific research. Their small size, short life span, and similarity to humans make them ideal for studying a wide range of topics, from genetics to human disease.
Transfection is a fundamental molecular biology technique used to introduce exogenous nucleic acids, such as DNA, RNA, or oligonucleotides, into eukaryotic cells to study gene function, protein expression, and cellular pathways. Unlike transduction, which employs viral vectors for gene delivery, transfection utilizes non-viral approaches, including lipid-based reagents, electroporation, and nanoparticle-mediated delivery, to facilitate gene transfer. This method plays a critical role in life sciences and pharmaceutical research, enabling functional genomics, drug discovery, and therapeutic development. Transfection can be categorized into transient transfection, where exogenous genetic material remains episomal and is expressed for a limited time, and stable transfection, where the introduced nucleic acid is integrated into the host genome, allowing for long-term expression. Advances in transfection technologies continue to enhance efficiency, specificity, and biocompatibility, expanding its applications in regenerative medicine, gene therapy, and precision medicine… Continue Reading