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Sep 1, 2010 (Vol. 30, No. 15)

Tackling Transfection's Complexity

Improved and New Reagents Facilitate Work with Difficult-to-Transfect Cells

  • Chemical Reagents

    Click Image To Enlarge +
    High-performance plasmid transfection: Primary human small epithelial cells were transfected using Mirus Bio’s TransIT-2020 and an EGFP expression plasmid (4:1 reagent-to-DNA ratio). Images were taken 24 hours post-transfection using a Zeiss axiovert inverted fluorescence microscope.

    “The trend in transfection, as we see it, is that it’s moving increasingly toward more hard-to-transfect cell lines,” said Laura Juckem, Ph.D., R&D group leader at Mirus Bio. “We are finding that researchers want to be in more physiological cell types, such as primary cells, which are more relevant to the disease or process that they are studying.”

    In response to this need, the company developed a reagent called TransIT-2020, which is designed to transfect cells typically recalcitrant to transfection such as primary cells.

    These transfection technologies are based on different platforms depending on the type of nucleic acid to be transfected. “We found through extensive testing of our compound libraries that there are different thresholds of performance within our reagents, and some are better suited for use with a particular nucleic acid,” said Dr. Juckem. For example, she added, “We have a totally different reagent for messenger RNA than for other nucleic acids. It is the only one that is commercially available to deliver messenger RNA in the presence of serum.”

    Dr. Juckem’s presentation at “Bioprocess International” later this month will focus on maximizing protein yield in suspension CHO cells. “Efficient transient transfection methods will allow researchers to produce high titers of proteins during preclinical drug discovery testing while the stable cell lines are under development,” Dr. Juckem remarked.



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