BacMam Expression System
Gene expression in mammalian cells
About BacMam
BacMam is a modified baculovirus vector which efficiently transduces nearly all mammalian cells, including primary cultures, standard cell lines, and iPSC derived cells. Researchers in life science and drug discovery have used BacMam for years to transiently deliver target receptors, genetically-encoded biosensors and other constructs in their cells of interest.
Montana Molecular’s biosensors, receptors, and other tools are packaged in BacMam because of its many benefits. Here we explore these benefits in an effort to explain why so many researchers are switching from plasmid transfection, stable cell lines and other viral vectors to using the BacMam system in their cells of choice.
- Consistent cell-to-cell, well-to-well, and day-to-day expression
- Robust expression in most mammalian cells – even iPSCs and primary cultures
- Viruses can easily be titrated for rapid assay optimization or mixed to express multiple genes in the same cells
- Simple workflow without extra incubation steps
Consistent, Titratable Expression
BacMam provides a very even level of expression from cell to cell and plate to plate, providing near stable-line consistency in a transient model. Expression can be easily titrated to a level that is suitable to the experiment and sensor being used. Receptors packaged in BacMam, for example, can be titrated to express at near endogenous levels. Experiments are more consistent, repeatable, and reliable.
DAG sensor expressing in HEK293T cells, comparing BacMam transduction (left) and plasmid transfection (right)
cAMP dose response curves at varying levels of GPCR expression
Expression in Difficult Cell Types and Primary Cultures
Our BacMam-packaged sensors have been used successfully in many primary and iPSC-derived cells including neurons, pancreatic islets, and cardiomyocytes. BacMam is a great choice for examining these interesting, disease-relevant cells because of its low toxicity and effective expression which can last for several days. Let us know if you have questions about your specific cell type, and see examples on our Publications page.
cADDis cAMP sensor expressing in Peri.4u iPSC derived peripheral neurons from Ncardia. Wide Field Fluorescence (left), Brightfield (right)
Testimonials
“The transfection efficiency of the BacMam is excellent in hard-to-transfect cell lines like Calu-3s (>30-40% vs <10% with lipofectamine) and we can even achieve 5-10% transduction in macrophages, which are nearly impossible to transfect with cationic lipids.”
Dr. Rob Lee
University of Pennsylvania
“Expression in human islets is pretty superb, after 18 hours of incubation. An excellent probe!”
Dr. Andrei Tarasov
Oxford Center for Diabetes
“Assay is working beautifully with cardiomyocytes, showing a strong and immediate response.”
Dr. Kit Werley
Harvard University / Q-State Biosciences
Available BacMam Vectors
Our off-the-shelf kits use one of two vectors: Big Sky BacMam or the Original (OG) BacMam vector
Big Sky BacMam
Big Sky BacMam is an improvement on our original vector, providing a robust increase in expression. We see a 5x boost in HEK293s, a 3x boost in CHO cells at 24 hours, and a 9x improvement in CHOs at 48 hours. Early adopters already report dramatic improvements in primary cultures. Our newest products, like cell painting tools and AKAP-targeted cADDis are provided in Big Sky, along with our cAMP, DAG, ER Stress, and Calcium biosensors.
Original (OG) BacMam
Unless otherwise noted, our products are mainly packaged in OG BacMam. We expect to be moving more products into Big Sky BacMam as we replenish our stocks. While not as highly expressing as Big Sky, it is still a fantastic vector and provides robust expression in most mammalian cell types. We see near 100% transduction efficiency in HEK293 cells, and our customers have published using products in cells ranging from primary neurons to pancreatic islets to macrophages.
Have specialized experimental needs? Learn more about our Evergreen and BacMammoth vectors for HDAC inhibitor-free expression and protein production!
Test Expression Efficiency
Test any of our vectors in your cells of choice with BacMam expressing red or green fluorescent proteins.
Interested in testing multiple BacMam versions or promoters in your cells? Reach out to our sales team for a quote.
Simple Workflow and Versatility
We also like BacMam for its ease of use. Simply add BacMam to your cells, incubate for 20-24 hours, and conduct your experiments. Incubation times of 48 hours can be helpful when expressing red fluorescent constructs. Separate viruses containing multiple constructs can be added at once, and an HDAC inhibitor boosts and maintains expression.
There are no extra incubation steps that you would have in a typical plasmid transfection and antibiotic can be present in the media. The same BacMam virus can also be used across a variety of cell types, no need to create multiple stable lines expressing the same receptor or construct.
Custom BacMam
Want to use BacMam to express genes that are not on our Product List? Need cell-specific promoters, point mutations, species orthologues, or other alterations to existing products? Montana offers BacMam production as a service to provide researchers with the most appropriate and precise tools for their experiments.
Get A Quote
References
BacMam: versatile gene delivery technology for GPCR assays: doi.org/10.1007/978-1-60327-317-6_14
Implementation of BacMam virus gene delivery technology in a drug discovery setting: doi.org/10.1016/j.drudis.2007.02.017
BacMam technology and its application to drug discovery: doi.org/10.1517/17460441.2.12.1669
BacMam recombinant baculoviruses in G protein-coupled receptor drug discovery: doi.org/10.1080/10606820490514969
New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories: dx.doi.org/10.1177%2F1087057115618608
Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli: dx.doi.org/10.1128/JVI.67.8.4566-4579.1993
