Custom BacMam
Express target genes in live cells with bespoke baculoviral vectors
Custom BacMam Production
Researchers in life science and drug discovery have used BacMam for years to transiently deliver target receptors other constructs in their cells of interest – an effective alternative to plasmid transfection or stable cell line development.
Montana Molecular provides full-service BacMam production – including DNA preperation, cloning, sequence confirmation, viral production, and qPCR to determine viral titer.
BacMam expression efficiency is greatly improved compared to plasmid transfection, enabling experiments in specialized, disease-relevant cell types. We see expression in 100% of HEK293T cells, and expression is extremely consistent from cell-to-cell, similar to stable cell lines. Researchers also use our BacMam tools in cells ranging from primary neurons to islets to macrophages and more.
In comparison to stable cell line development, a key advantage of BacMam is the flexibility to express genes across cell lines, iPSCs, and primary cultures using the same reagent. Expression levels are easily titratable, allowing rapid assay optimization. BacMam production is faster and lower cost than stable cell lines, enabling groups to screen and test species orthologues or mutations in their target proteins.
Our scientists bring years of expertise in gene expression and molecular biology, work with us to develop the right tool for your in vitro assays!
Consistent cell-cell expression of our fluorescent DAG sensor in HEK293T cells using BacMam
Green Fluorescent cADDis cAMP sensor expressed in Primary Striatal Neurons with BacMam
Customizable
Work with Montana Molecular scientists to develop your ideal research tool. Choose cell-specific promoters, enable cre-dependent expression, add genetic tags or fluorescent protein biomarkers, knock out genes with CRISPR/CAS systems and more! Our selection of next-generation BacMam vectors address specialized experimental needs, including HDAC inhibitor-free expression.
Flexible
Robustly express your target gene across standard cell lines, primary cultures, and iPSCs with a single reagent. Expression levels are easily tunable by adjusting the amount of virus added to your cells, facilitating rapid assay optimization and enabling expression at near endogenous levels. Multiple BacMam viruses can be added together to co-express genes in the same cell population.
Time and Cost Savings
Rapidly screen and test multiple species orthologues or mutations with BacMam. Production of high-titer P3 virus takes ~6 weeks and multiple viruses can be produced in parallel, prioritized according to your interests. We archive P2 for fast, cost-effective scale-up if additional virus from hits or targets is needed.
Large Payload
Express large genes or constitutively express several genes, taking advantage of BacMam’s >34kb genetic payload.
Next-generation BacMam vectors
Find the best BacMam vector for your experiments
Big Sky BacMam
Yields the best transduction efficiency in the widest range of cells. We see a 4-5x boost in expression in HEK and CHO cells, and our customers have reported even better results in primary cultures.
Evergreen BacMam
Expression in human or primate cells without the need for HDAC inhibitors. Ideal for experiments with cells that are particularly sensitive to HDAC inhibitors like iPSCs and certain primary cultures.
BacMammoth System
A two-vector system yielding high levels of protein. One vector delivers the target gene and a “booster” virus enhances protein production. A great tool for generating protein for structural studies.
Test Expression Efficiency in your Cells
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 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 for Original, Big Sky, and BacMammoth vectors.
Transduce cells in suspension or in adherent cultures. Large batches of cells may be transduced and added directly to compounds, or frozen for later use. BacMam is an extremely versatile tool, and the protocol can easily be tailored to suit your experiments and instrumentation.
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.
Gene Knockout with BacMam
Use a two-vector CRISPR system to knockout genes with BacMam. One vector expresses the Cas9 enzyme, while the other delivers a custom CRISPR guide. Knock out genes in standard cell lines, iPSCs, or primary cultures.
Our example figure shows data from HEK293 cells expressing our cADDis cAMP sensor alone, with Cas9, or with Cas9 and a CRISPR guide for the Gαs protein. Note that the cells expressing the Cas9 and guide do not respond to isoproterenol stimulation of endogenous Beta-2 adrenergic receptors, but still respond to adenylyl cyclase activation by forskolin.
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References
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
