Demonstrated in Science¹, our exclusively licensed F.A.S.T.™ (Flexible Accelerated STOP TetO) technology is a versatile tool where a single targeting event yields a multi-purpose mouse line with the potential for five or more controllable functionalities from the same locus, just using mating.² These functionalities can include global gene knockout, tissue/time-specific gene rescue, ectopic gene expression, inducible/reversible gene overexpression, and inducible/reversible gene knockdown.
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.
1) 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.
2) 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).
3) 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.
1) 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. Science 338(6103): 128-132.
2) 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.
1) Tet Systems: Principles and Components
2) Tet Systems: Home Page
3) Schönig K, Bujard H, Gossen M. 2010. The power of reversibility: regulating gene activities via tetracycline-controlled transcription. Methods Enzymol 477: 429-453.
4) 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.