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AfCS Technology Development

The AfCS laboratories are developing several key technologies which will provide an important experimental basis for our ongoing research efforts. These efforts are focused in the following areas:

  • The development of optimized approaches for RNA interference in AfCS model cell types. We have concentrated on viral vector-based strategies for shRNA expression, with an emphasis on the development of reliable platforms for conditional shRNA expression. We are also routinely using a protocol we optimized for transient transfection of synthesized siRNAs, as this allows us to quickly identify key targets for knockdown (KD) that can be prioritized for viral-based studies.
  • Application of expression techniques to cDNAs. Both retroviral and lentiviral platforms are being used to identify optimal strategies for both constitutive and inducible cDNA expression.
  • The development and optimization of FRET and BRET-based biosensors for real-time measurement of key signaling intermediates in live cells.
  • Establishment of an experimental platform for a primary cell counterpart to RAW264.7 cells. This involves the differentiation of primary macrophages from bone marrow-derived hematopoietic precursors and the assay and experimental manipulation of these cells.

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