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Category Archives: Transfection

Nanoparticle In Vivo Transfection Reagent (Altogen)

July 27, 2021 – Altogen Biosystems® is a biotechnology manufacturing company that specializes in the development and testing of optimized cell transfection reagents and kits for both in vitro and in vivo delivery. In 2018 Altogen Biosystems launched the AltoFect transfection reagent formulated for difficult-to-transfect cells (like B-cells, T-cells), and primary cell cultures that are known to be resistant to most typical transfection technologies. Company offers over 120 cell-line specific transfection kits, optimized electroporation products, and  in vivo transfection reagents that are used for tissue-targeted delivery of negatively charged biomolecules (siRNA/miRNA, mRNA, plasmid DNA, small proteins, etc).

Altogen Biosystems announced that its Nanoparticle-based In VivoTransfection Reagentwas used for effective overcoming blood-brain barrier and successful transfection of glioblastoma tumor cells with both plasmid DNA and siRNA molecules. Altogen Labs mouse orthotopic PDX glioblastoma xenograft model was used for the study. This xenograft model reproduce glioblastoma histologic complexity and many biological behaviors (angiogenesis, brain invasion, and resistance to therapy). Altogen Labs research study demonstrated functional cargo delivery of both siRNA and DNA (3.72Kb) through the blood–brain barrier, as well as high transfection efficiency and reproducibility. Nanoparticle-based In VivoTransfection Reagent is available as reagent-grade that is commonly used for preclinical research studies, as well as GMP grade product manufactured for clinical research applications. Altogen’s nanoparticle in vivo transfection reagent was referenced in over 40 research publications for effective in vivo delivery of biomolecules, however Altogen scientists still had to develop modified in vivo transfection protocol to enable brain-targeted delivery.

About Altogen Biosystems

Altogen Biosystems is a life sciences products manufacturer that offers a spectrum of transfection kits and electroporation buffers that are optimized for cancer cell lines and primary cell types. Altogen’s in vivo transfection reagents enable tissue-targeted delivery of any negatively charged moleculess. Scientists at Altogen Biosystems utilize chemical engineering and cell biology expertise to develop novel efficient and safe transfection products to assist scientists in revolutionary cancer and drug development research.

Altogen Biosystems
848 Rainbow Blvd #823
Las Vegas, NV, 89107 USA
Phone: (702) 349-6103
Fax: (702) 989-0841
Email: corporate@altogen.com

Crossing Blood-Brain Barrier and In Vivo Delivery of Negatively Charged Biomolecules into Glioblastoma Enabled by Altogen's Nanoparticle In Vivo Transfection Reagent

LAS VEGAS, NEVADA – July 19, 2021 – Altogen Biosystems, leading manufacturer of in vivo transfection products, announced today that Altogen’s nanoparticle-based in vivoTransfection reagent (catalog #5031) was demonstrated to be effective for glioblastoma-targeted co-delivery of plasmid DNA (3.7 kb) and chemically modified siRNA. This transfection reagent was used in over 30 research publications, demonstrating highly… Continue Reading

Citations and References: Altogen® Transfection Kits

Citations and References: ALTOGEN® Transfection Reagents Nature 2008 454(7203):523-7. Innate immunity induced by composition-dependent RIG-I …Saito et al [PDF] Nature Medicine 2016 22(10):1131–1139. A Long Non-Coding RNA Defines an Epigenetic Checkpoint … Wang et al [PDF] Cell Rep 2018 22(2):523. The Foxo1-Inducible Transcriptional Repressor Zfp125 … Gustavo et al [PDF] Molecular Therapy 2018. Liposome Lipid-Based… Continue Reading

cGMP-grade Nanoparticle Transfection Reagent for In Vivo Delivery

Altogen Biosystems is a manufacturer of life science research products that includes in vivo transfection reagents designed for tissue-targeted delivery of biomolecules. Altogen’s Nanoparticle In Vivo Transfection Reagent was launched in 2008 and used in over 30 research publications demonstrating highly efficient delivery of cargo nucleic acids (siRNA, microRNA, mRNA, plasmid DNA, and small proteins)… Continue Reading

CRISPR/Cas9 Transfection Optimization

Efficient delivery of the CRISPR/Cas9 protein complex can be achieved efficiently through the use of cationic lipid-based transfection reagents. Cells may undergo electroporation to increase the probability of successful transfection. Once inside a target cell, the CRISPR/Cas9 protein complex requires no additional stimulation to start modifying the cell’s genome. Genome modification will occur within one… Continue Reading

Forward Transfection or Reverse Transfection?

Forward and reverse transfection protocols each have their significant uses in research.  The main protocol difference between forward and reverse transfection is whether or not the cells are plated the day before transfection (as in forward transfection) or seeded at the same time of the transfection.  Forward transfection is commonly used in situations where the… Continue Reading

AltoFect v2.0 second generation transfection reagent

AltoFect v2.0, the second generation transfection reagent for primary cell types and hard-to-transfect cell lines. AltoFect transfection reagent exhibits up to 85% transfection efficiency in difficult-to-transfect cells such as T-cells, B-cells, and primary cell cultures. The advance has opened up the path to reliable in vitro testing of realistic responses from cells that were previously… Continue Reading

Plasmid DNA and siRNA transfection protocol optimization

There are several experimental parameters that affect transfection efficiency and associated cell viability for siRNA and plasmid DNA transfection experiments: Transfection protocol (method and conditions) Health of cultured cells Purity and concentration of pDNA and/or siRNA Controls Transfection protocol Optimized transfection protocols are available for over 100 cancer cell lines and primary cell types (see… Continue Reading

Stable transfection reagents and techniques

Stable transfection, sometimes called permanent transfection, is the integration of plasmid DNA into the chromosome of cancer cell’s DNA.  The creation of a stable cell line enables the researcher to analyze long term effects of the introduced gene.  Downstream studies incorporating the stable cell line includes overexpression of the gene insert for protein production, cell… Continue Reading

How much antibiotic required for stable cell selection?

Stable cell lines are a crucial laboratory tool that over-expresses a gene of interest in order to study gene functions, screen experimental drugs or produce therapeutic proteins (i.e. recombinant antibodies).  The cell lines will divide and continue to express the inserted transgene.  Briefly, exogenous plasmid DNA is transfected into a host cell line, which is… Continue Reading

Transfection optimization

The most vital aspect of a transfection is ensuring all conditions are optimized, including transfection reagent volume, oligo concentration, cell viability and passage number, lack of negative control activity, activity of positive control and calibrated incubator temperature, humidity and percentage of CO2.  Even if all these parameters are correctly addressed, transfection efficiency must be determined… Continue Reading

Importance of cell passage number and cell confluency for efficient transfection

The viability, confluency and passage number are all vital parameters for a successful transfection.  Here are helpful hints for each of these aspects: Viability Viability is the percentage of living cells in a suspension Determined using trypan blue exclusion Cells are considered to be healthy if viability is greater than 90% Confluency Confluency is the… Continue Reading

Quantitate in vitro anti-proliferation experiments using a metabolic assay (Alamarblue, MTT) vs measuring protein abundance (Sulforhodamine B)

The need to measure cell proliferation effects is vast, including testing the effects of growth factors, novel pharmacological agents, cytotoxicity assessment or investigating cell activation.  Cell proliferation assays utilize a stain or substrate to make a correlation between the readout and the number of remaining cells post-treatment.  However, the means of assessing cell number varies… Continue Reading

What is the benefit of using an electroporation buffer

The process of electroporation exposes the cells to a high-voltage pulse of electricity to disrupt the phospholipid bilayer of the cell membrane causing the formation of temporary pores.  Any charged molecules (e.g. RNA, DNA) are forced into the cells thru the pores. Electroporation buffers are formulations that mimic cellular cytoplasm composition; thus, enhancing pore resealing… Continue Reading

Difference between technical and biological replicates

The basic definitions of technical and biological replicates are as follows: Technical replicates: a test performed on the same sample multiple times; i.e., if there are triplicate non-treated samples, a technical replicate would be testing sample #1 of the non-treated multiple times Biological replicates: a test performed on biologically distinct samples representing an identical time… Continue Reading

What is the best method to detach cultured cells?

Cultured adherent cells routinely need to be detached and collected for counting or passaging.  Detaching cells can be accomplished by either mechanical or enzymatic methods. Mechanical: cell scraping is a good option for cells that are sensitive to trypsin but can cause damage to cells; also can be used when collecting cellular components for western… Continue Reading

What transfection controls do I need to include in my experiment?

Scientists understand the importance of controls in experiment. There are at least three transfection controls that should be included on every transfection plate: a positive control, negative control and non-treated control. Positive Controls Transfecting a positive control ensures that the system being utilized is working and the delivery conditions are optimal.  The results of the… Continue Reading

What is the best antibiotic to use for stable cell selection in mammalian cells?

There is a long list of antibiotics available to researchers when applying selective pressure in the creation of a stable cell line. The choices include zeocin, hygromycin, blasticidin, puromycin and geneticin (G418). Researchers use different antibiotics due to cost or availability in their lab. However, in reference to using antibiotics for the creation of a… Continue Reading

How do I increase transfection efficiency?

Transfection is considered as a major laboratory method to integrate protein, RNA and DNA molecules into tissues and cells. Delivery of plasmid DNA molecules containing gene inserts, messenger RNA and small interfering RNA molecules that regulate gene expression (i.e. microRNA, siRNA) into the primary cells and cancer cell lines have been extensively utilized by scientists.… Continue Reading