Flexible Accelerated STOP Tetracycline Operator-knockin (FAST): a versatile and efficient new gene modulating system.
We created the Flexible Accelerated STOP Tetracycline Operator (tetO)-knockin (FAST) system, an efficient method for manipulating gene expression in vivo to rapidly screen animal models of disease. A single gene targeting event yields two distinct knockin mice-STOP-tetO and tetO knockin-that permit generation of multiple strains with variable expression patterns: 1) knockout, 2) Cre-mediated rescue, 3) tetracycline-controlled transcriptional activator (tTA)-mediated misexpression, 4) tetracycline-controlled transcriptional activator (tTA)-mediated overexpression, and 5) tetracycline-controlled transcriptional silencer (tTS)-mediated conditional knockout/knockdown. Using the FAST system, multiple gain-of-function and loss-of-function strains can therefore be generated on a time scale not previously achievable. These strains can then be screened for clinically relevant abnormalities. We demonstrate the flexibility and broad applicability of the FAST system by targeting several genes encoding proteins implicated in neuropsychiatric disorders: Mlc1, neuroligin 3, the serotonin 1A receptor, and the serotonin 1B receptor.
Tanaka
Kenji F
KF
Division of Neurobiology and Bioinformatics, National Institute for Physiological Sciences, Okazaki Aichi, Japan.
Ahmari
Susanne E
SE
Leonardo
E David
ED
Richardson-Jones
Jesse W
JW
Budreck
Elaine C
EC
Scheiffele
Peter
P
Sugio
Shouta
S
Inamura
Naoko
N
Ikenaka
Kazuhiro
K
Hen
René
R
D000818:Animals; D030801:Animals, Genetically Modified; D001253:Astrocytes / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D015870:Gene Expression / Q000235:genetics / Q000502:physiology; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D001523:Mental Disorders / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009422:Nervous System Diseases / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*
D001426:Bacterial Proteins; D002352:Carrier Proteins; D012333:RNA, Messenger; C083139:Tet O resistance protein, Bacteria; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D013752:Tetracycline