B6;C3-Tg(NEFH-tTA)8Vle Tg(tetO-TARDBP*)4Vle/J

Stock number: 028412
Product name: rNLS8 (or NEFH-hTDP-43ΔNLS)
Strain synonyms: rNLS8, NEFH-hTDP-43DeltaNLS
Research areas: Neurobiology Research, Research Tools

Description

The Jackson Laboratory will not distribute double transgenic mice. Instead, we distribute the individual transgenic lines - mice having either the NEFH-tTA transgene (see Stock No. 025397) or the tetO-hTDP-43-ΔNLS transgene (see Stock No. 014650). This regulatable NLS (rNLS8) double transgenic line recapitulates cytoplasmic TDP-43 pathology reminiscent of amyotrophic lateral sclerosis/frontotemporal lobar degeneration in brain and spinal cord, accompanied by a progressive neurodegenerative ALS-like phenotype. The phenotype is controllable/reversible by the tetracycline analog, doxycycline (dox).

Users should be aware that mice harboring both transgenic alleles will exhibit the lethal characteristics of amyotrophic lateral sclerosis (ALS)/frontotemporal lobar degeneration and must be provided tetracycline or a suitable analog such as doxycycline for longer term maintenance.

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Detailed description

These mice are useful in studying amyotrophic lateral sclerosis (ALS) / frontotemporal lobar degeneration (FTLD). The phenotype of these mice is expected to be as described in Walker et al. 2015 Acta Neuropathol 130:643; details below.

Stock No. 028412 is the regulatable NLS (rNLS8) double transgenic line created by breeding NEFH-tTA line 8 (Stock No. 025397) with TRE-promoter-driven cytoplasmic-insoluble human TDP-43ΔNLS transgenic mice (tetO-hTDP-43-ΔNLS line 4 ; Stock No. 014650). On a mixed C57BL/6J x C3HeJ F1 genetic background, mice hemizygous for NEFH-tTA and hemizygous for tetO-hTDP-43-ΔNLS exhibit lethal characteristics of amyotrophic lateral sclerosis (ALS) / frontotemporal lobar degeneration (FTLD) when maintained in the absence of the tetracycline analog, doxycycline (dox). Specifically, they exhibit widespread, high levels of hTDP-43ΔNLS expression in neurons of both the brain and spinal cord (SC) as early as 1 week off-dox. The brain/SC combination of hTDP-43-ΔNLS expression accounts for the spinal motor neuron loss and muscle denervation; resulting in a rapid, robust and progressive neurodegenerative ALS-like phenotype (including motor deficits, denervation of neuromuscular junctions, motor neuron loss and premature death at ~8-20 weeks). Upon administration of dox, the observed TDP-43 pathology and functional deficits were found to be largely reversible, even after neurodegeneration was underway (~8 weeks of hTDP-43-ΔNLS expression).

Stock No. 028413 is the regulatable control double transgenic line created by breeding NEFH-tTA line 8 (Stock No. 025397) with TRE-promoter-driven human TDP-43 wildtype transgenic mice (tetO-hTDP-43-WT line 12 ; Stock No. 016841). On a mixed C57BL/6J x C3HeJ F1 genetic background, mice hemizygous for NEFH-tTA and hemizygous for tetO-hTDP-43-WT exhibit broad nuclear hTDP-43-WT expression in the brain and spinal cord, without the formation of cytoplasmic TDP-43 pathology.

Importantly, the donating investigator reports that the human NEFH promoter in NEFH-tTA line 8 drives slightly higher levels of tTA expression in the cortex compared to their experiences with the mouse Camk2a promoter used in Camk2a-tTA mice (greater than ~10-fold higher than endogenous NEFH versus ~8-fold higher than endogenous CAMK2A). In addition, the human NEFH promoter directs tTA expression in spinal cord (whereas the mouse Camk2a promoter did not).

Development

Stock No. 028412 is the regulatable NLS (rNLS8) double transgenic line created by breeding NEFH-tTA line 8 (Stock No. 025397) with TRE-promoter-driven cytoplasmic-insoluble human TDP-43ΔNLS transgenic mice (tetO-hTDP-43-ΔNLS line 4 ; Stock No. 014650). Both transgenes were designed in the laboratory of Dr. Virginia Man-Yee Lee (University of Pennsylvania).

For NEFH-tTA line 8 (Stock No. 025397), the donating investigator reported multiple transgene copies are likely as the expression level is 9-10-fold greater than endogenous levels [September 2015]. In 2020, McMillan et al. 2020 Acta Neuropathol Commun 8:200 reported that mice obtained from The Jackson Laboratory Stock No. 025397 have the NEFH-tTA transgene inserted into chromosome 12 [Chr12:6,917,892-6,917,912] without any disruption of gene regions, but did not indicate any transgene copy number information.

Allele

Symbol: Tg(tetO-TARDBP*)4Vle
Name: transgene insertion 4, Virginia M Y Lee
MGI accession ID: MGI:5000261
Generation method: Transgenic
Attributes: Inducible; Inserted expressed sequence; Humanized sequence
Synonyms: hTDP-43deltaNLS4
Marker symbol: Tg(tetO-TARDBP*)4Vle
Marker name: transgene insertion 4, Virginia M Y Lee
Chromosome: UN
Strain of origin: (C57BL/6J x C3H/HeJ)F1
Expressed genes: TARDBP,TAR DNA binding protein,human
Site of expression: When bred to mice expressing tetracycline transactivator (tTA), tetracycline or its analog doxycycline (DOX) administration results in the expression of mutant human TARDBP, TAR DNA binding protein, hTDP-43-deltaNLS in transactivator expressing tissues of the offspring.
Molecular note: The tetracycline responsive promoter drives inducible expression of the human cDNA lacking the nuclear localization signal. Two lines were generated (4 and 19).

Allele

Symbol: Tg(NEFH-tTA)8Vle
Name: transgene insertion 8, Virginia M-Y Lee
MGI accession ID: MGI:5693898
Generation method: Transgenic
Attributes: Transactivator
Synonyms: NEFH-tTA
Marker symbol: Tg(NEFH-tTA)8Vle
Marker name: transgene insertion 8, Virginia M-Y Lee
Chromosome: 12
Strain of origin: (C57BL/6J x C3H/HeJ)F1
Expressed genes: tTA,tetracycline-controlled transactivator,E. coli
Site of expression: tTA is expressed in developmental late stage of the cortex, cerebellum and hippocampus, with expression also observed in olfactory bulb and in the rest of the brain.
Molecular note: The construct consists of ~18 kbp of the human neurofilament heavy polypeptide promoter (NEFH; from BAC RP11-91J21) upstream of the tetracycline-regulated transactivator gene (tTA or "Tet-Off"), followed by a polyA signal. Founder line 8 has its highest tTA expression in developmental late stage of the cortex, cerebellum and hippocampus, with expression also observed in olfactory bulb and in the rest of the brain.