mouse thy1 2 promoter  (Addgene inc)

Journal: Cell reports

Article Title: Distinct Mechanisms of Over-Representation of Landmarks and Rewards in the Hippocampus

doi: 10.1016/j.celrep.2020.107864

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse Thy1.2 promoter , Feng et al., 2000 , RRID: Addgene_20736.

Techniques: Recombinant, Software, Imaging, Modification

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Mutant PFN1 causes ALS phenotypes and progressive motor neuron degeneration in mice by a gain of toxicity.

doi: 10.1073/pnas.1605964113

Figure Lengend Snippet: Fig. 1. Exogenous PFN1 is predominantly expressed in the CNS within trans- genic lines. (A) Transgene constructs expressed PFN1 cDNA with an N-terminal V5 epitope tag. Transgenic lines used the mouse prion (Prp) promoter to drive expression of either wild-type (Prp-PFN1WT) or an ALS-associated mutant (Prp- PFN1C71G) PFN1. An additional mouse line used the human Thy1.2 promoter to drive expression of mutant PFN1 (Thy1.2-PFN1C71G). (B) Western blots demon- strate that four transgenic lines (two wild-type, two mutant) express the transgenes (upper band) predominantly in the CNS. Lanes represent different organs: 1, forebrain; 2, cerebellum; 3, brainstem; 4, cervical spinal cord; 5, lumbar spinal cord; 6, heart; 7, lung; 8, liver; 9, spleen; 10, kidney; 11, muscle. nTg, nontransgenic mice. (C) Western blots for PFN1 demonstrate the relative expression of exogenous PFN1 in the spinal cord after intercrossing the two mutant transgenic lines. endo, endogenous PFN1; Tg, V5-PFN1 transgene; 1, Thy1.2-PFN1C71G/C71G/Prp-PFN1C71G triple transgenic mice; 2, Thy1.2-PFN1C71G/C71G homozygous mice; 3, Thy1.2-PFN1C71G/Prp-PFN1C71G

Article Snippet: The cDNAs encoding human PFN1C71G and PFN1WT with N-terminal V5 tag were cloned into the XhoI site of the MoPrp.Xho plasmid (ATCC#JHU-2) or mouse thy1.2 plasmid (gift from Joshua Sanes, Harvard University, Cambridge, MA; Addgene plasmid 20736).

Techniques: Construct, Transgenic Assay, Expressing, Mutagenesis, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Mutant PFN1 causes ALS phenotypes and progressive motor neuron degeneration in mice by a gain of toxicity.

doi: 10.1073/pnas.1605964113

Figure Lengend Snippet: Fig. 2. Transgenic mice expressing mutant but not WT PFN1 show progressive loss of motor capabilities. (A) Photo of a Thy1.2-PFN1C71G transgenic at ∼400 d old displaying ppar (Movie S1). (B) The onset age of paralysis of mutant PFN1 mice are shown: 1, Thy1.2-PFN1C71G mice; 2, Thy1.2-PFN1C71G/C71G mice; 3, Thy1.2- PFN1C71G/Prp-PFN1C71G mice; 4, Thy1.2-PFN1C71G/C71G/Prp-PFN1C71G mice. The rank order of the ages correlated inversely with the expression of the mutant PFN1 protein (Fig. 1 C and D). The plots represent data from both genders, as this did not appear to contribute to disease onset or progression (Fig. S2A). (C) Rotorod tests demonstrated stable performances in the nTg and PFN1WT mice but a progressive decline in the PFN1C71G mice after 4 mo of age. All time points represent averages of 5–28 mice, 4–21 mice, and 10–25 mice for the nTg, PFN1WT, and PFN1C71G groups, respectively, with the exception of the end point for PFN1C71G (n = 2). Error bars represent the SE. (D) PFN1C71G mice display a progressive decline in home cage vertical behavior (rearing, jumping, hanging, climbing, and coming down). Vertical time points represent averages from 8 to 10 mice, 7–10 mice, and 8–10 for the nTg, PFN1WT, and PFN1C71G groups, respectively. Error bars are SE. (E) The peak body weight of PFN1C71G mice is reached between age 4 and 6 mo and declined afterward. In contrast, the nTg and PFN1WT mice progressively increased body weight beyond this time period. Female mice display a similar pattern of weight loss (Fig. S2B). Time points are averages of 11–30 mice, 3–18 mice, and 9–21 mice for the nTg, PFN1WT, and PFN1C71G groups, respectively. Error bars represent the SD. (F) Grip strength tests demonstrated stable performances in the nTg and PFN1WT mice but a progressive decline in PFN1C71G mice after peaking at 3 mo of age. Time points are averages of 8–25 mice, 6–15 mice, and 11–32 mice for the nTg, PFN1WT, and PFN1C71G groups, respectively. The color representation of mouse genotypes in C–F is shown in D. Error bars represent the SE.

Article Snippet: The cDNAs encoding human PFN1C71G and PFN1WT with N-terminal V5 tag were cloned into the XhoI site of the MoPrp.Xho plasmid (ATCC#JHU-2) or mouse thy1.2 plasmid (gift from Joshua Sanes, Harvard University, Cambridge, MA; Addgene plasmid 20736).

Techniques: Transgenic Assay, Expressing, Mutagenesis