The F.A.S.T.™ (Flexible Accelerated STOP TetO) technology by ingenious targeting laboratory offers researchers a versatile and powerful tool for generating multi-purpose mouse lines with controllable functionalities, all from a single targeting event. Demonstrated in Science¹, this technology allows for five or more controllable functionalities from the same locus, simply through mating², streamlining the mouse model generation process. Options include global gene knockout, tissue/time-specific gene rescue, ectopic gene expression, inducible/reversible gene overexpression, and inducible/reversible gene knockdown.
These options can be applied to your gene of interest using the F.A.S.T.™ cassette, providing flexibility and versatility for your research endeavors. By incorporating this technology, researchers can achieve precise control over gene expression levels and patterns, enhancing the adaptability and efficiency of their experimental designs.
“The project was very well managed…in fact, using iTL validated my decision to not try and do this in my own lab. It would have been a catastrophe… (My project manager) was very helpful, always getting back to us in time and explaining every step of the project. I would be glad to serve as a reference for iTL and its staff.”– Claus Fimmel, MD Loyola University Medical Center
Depicted in the schematic below, Tanaka KF et al showed that the F.A.S.T.™ system achieves a spectrum of controllable expression levels from the Mlc1 gene, thereby streamlining the mouse model generation process.
As a knockout first, the Mlc1 gene function was rescued by mating the initial mouse line to a tissue specific Cre line of choice. Alternatively, gene expression can be induced through the F.A.S.T.™ cassette. A tetracycline transactivator (tTA) line was used to produce an ectopic gene expression model, and a tissue-specific over-expression model. A tetracycline trans-silencer line was used to generate a tissue-specific conditional knockdown/knockout.
Salvatierra J, Diaz-Bustamante M, Meixiong J, Tierney E, Dong X, Bosmans F. 2018. A disease mutation reveals a role for NaV1.9 in acute itch. J Clin Invest 128(12): 5434-5447.
Wallace CH, Wu BX, Salem M, Ansa-Addo EA, Metelli A, Sun S, Gilkeson G, Shlomchik MJ, Liu B, Li Z. 2018. B lymphocytes confer immune tolerance via cell surface GARP-TGF-β complex. JCI Insight 3(7).
Wu BX, Li A, Lei L, Kaneko S, Wallace C, Li X, Li Z. 2017. Glycoprotein A repetitions predominant (GARP) positively regulates transforming growth factor (TGF) β3 and is essential for mouse palatogenesis. J Biol Chem 292(44): 18091-18097.
Baudouin SJ, Gaudias J, Gerharz S, Hatstatt L, Zhou K, Punnakkal P, Tanaka KF, Spooren W, Hen R, De Zeeuw CI, Vogt K, Scheiffele P. 2012. Shared synaptic pathophysiology in syndromic and nonsyndromic rodent models of autism.Science338(6103): 128-132.
Tanaka KF, Ahmari SE, Leonardo ED, Richardson-Jones JW, Budreck EC, Scheiffele P, Sugio S, Inamura N, Ikenaka K, Hen R. 2010. Flexible Accelerated STOP Tetracycline Operator-knockin (FAST): a versatile and efficient new gene modulating system.Biol Psychiatry 67(8): 770-773.
Tet Systems: Principles and Components
Tet Systems: Home Page
Schönig K, Bujard H, Gossen M. 2010. The power of reversibility: regulating gene activities via tetracycline-controlled transcription.Methods Enzymol 477: 429-453.
Stieger K, Belbellaa B, Le Guiner C, Moullier P, Rolling F. 2009. In vivo gene regulation using tetracycline-regulatable systems.Adv Drug Deliv Rev 61(7-8): 527-541.
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