Drosophila TREM2 and TYROBP Models

Flies were maintained in standard cornmeal media at 25 °C. Complementary DNA (cDNA) encoding the full length of TREM2 (NM_018965, RC221132) and TYROBP (NM_198125, RC203771) with Myc-DDK tag were obtained from OriGene Technologies, Inc. These constructs were subcloned into a pJFRC19-13XLexAop2 vector (Addgene #26224). TREM2R47H mutation was introduced by using site-directed mutagenesis kit (Takara Bio Inc.). Transgenic flies were generated by PhiC31 integrase-mediated transgenesis systems (Best Gene Inc.). Transgenic fly lines carrying UAS-Aβ42 and UAS-tau were previously described [43 (link)–45 (link)]. Repo-LexA (#67096), Elav-GAL4 (#458), GMR-GAL4 (#1104), UAS-para RNAi (#31626), and UAS-mcherry RNAi (#35785) were obtained from the Bloomington Stock Center. For RNA sequencing (RNA-seq), around seven-day-old male flies were used. All experiments were performed using age-matched male flies.

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Sekiya M., Wang M., Fujisaki N., Sakakibara Y., Quan X., Ehrlich M.E., De Jager P.L., Bennett D.A., Schadt E.E., Gandy S., Ando K., Zhang B, & Iijima K.M. (2018). Integrated biology approach reveals molecular and pathological interactions among Alzheimer’s Aβ42, Tau, TREM2, and TYROBP in Drosophila models. Genome Medicine, 10, 26.

Publication 2018

Myc-DDK tag site-directed mutagenesis kit PhiC31 integrase-mediated transgenesis systems Elav-GAL4 UAS-mcherry RNAi

Cdna Flies Gene Integrase Male Mutation Rnai Site directed mutagenesis Transgenic Trem2 Vector

Corresponding Organization :

Other organizations : National Center for Geriatrics and Gerontology, Icahn School of Medicine at Mount Sinai, The Neurological Institute, Columbia University Irving Medical Center, Rush University Medical Center, Tokyo Metropolitan University, Nagoya City University

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Variable analysis

independent variables

TREM2 (NM_018965, RC221132) and TYROBP (NM_198125, RC203771) with Myc-DDK tag
TREM2R47H mutation

dependent variables

Not explicitly mentioned

control variables

Flies were maintained in standard cornmeal media at 25 °C
Transgenic fly lines carrying UAS-Aβ42 and UAS-tau
Repo-LexA (#67096), Elav-GAL4 (#458), GMR-GAL4 (#1104), UAS-para RNAi (#31626), and UAS-mcherry RNAi (#35785) from the Bloomington Stock Center

controls

Positive control: Transgenic fly lines carrying UAS-Aβ42 and UAS-tau
Negative control: Not explicitly mentioned

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