RGD Reference Report - A novel vector for transgenesis in the rat CNS. - Rat Genome Database

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A novel vector for transgenesis in the rat CNS.

Authors: Lopez, T Peter  Giles, Kurt  Dugger, Brittany N  Oehler, Abby  Condello, Carlo  Krejciova, Zuzana  Castaneda, Julian A  Carlson, George A  Prusiner, Stanley B 
Citation: Lopez TP, etal., Acta Neuropathol Commun. 2017 Nov 21;5(1):84. doi: 10.1186/s40478-017-0484-y.
RGD ID: 15045596
Pubmed: PMID:29157304   (View Abstract at PubMed)
PMCID: PMC5697436   (View Article at PubMed Central)
DOI: DOI:10.1186/s40478-017-0484-y   (Journal Full-text)

The larger brain of the rat enables a much greater repertoire of complex behaviors than mice, likely making rats preferential for investigating neurodegeneration. Because molecular tools for specific expression of transgenes in the rat brain are sparse, we chose Prnp encoding the prion protein (PrP) to develop a novel vector to drive transgene expression in the rat brain. We compared the rat Prnp sequence with mouse and Syrian hamster Prnp sequences, identifying conserved genetic elements and hypothesizing that these elements would be able to drive neuronal transgene expression. We investigated this by generating a vector termed RaPrnp that encompasses portions of the rat Prnp gene. Importantly, we replaced the rat Prnp open reading frame (ORF) with a cloning site for rapid and seamless In-Fusion cloning. To validate the in vivo neuronal specificity of the RaPrnp vector in rats, we generated stable RaPrnp-LacZ/enhanced green fluorescent protein (EGFP) transgenic (Tg) rat lines, which led to robust LacZ activity and high EGFP fluorescence in the central nervous system of embryos and adult animals. Next, we restored the rat Prnp ORF and generated multiple Tg(RaPrnp-PrP) lines, demonstrating that overexpression of Prnp accelerates the onset of scrapie. While the incubation time in wild-type (WT) rats was 175 ± 3 days post inoculation (dpi), one line, Tg2919, overexpressed RaPrPC at 4.4-fold and exhibited a reduced incubation time of 149 ± 2 dpi. The second line, Tg2922, overexpressed RaPrPC at 9.7-fold compared with WT animals and had an incubation time of 112 ± 0 dpi. Tg2922 rats inoculated with rat RML showed extensive vacuolation of the brainstem in contrast to WT and Tg2919 animals in which vacuolation was most prominent in the hippocampus and striatum as well as the motor and sensory cortices. It is possible that construction of Tg rats with modified phenotypes will prove more advantageous than mice for neurodegeneration studies.



RGD Manual Disease Annotations    Click to see Annotation Detail View

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
PRNPHumanscrapie onsetISOPrnp (Rattus norvegicus) RGD 
PrnpRatscrapie onsetIMP  RGD 
PrnpMousescrapie onsetISOPrnp (Rattus norvegicus) RGD 
SD-Tg(Prnp-Prnp)2919Ratscrapie inducedIMPprion RGD 
SD-Tg(Prnp-Prnp)2922Ratscrapie inducedIMPprion RGD 

Objects Annotated

Genes (Rattus norvegicus)
Prnp  (prion protein)

Genes (Mus musculus)
Prnp  (prion protein)

Genes (Homo sapiens)
PRNP  (prion protein (Kanno blood group))

Objects referenced in this article
Strain SD-Tg(Prnp-LacZ/EGFP)12084 null Rattus norvegicus
Strain SD-Tg(Prnp-LacZ/EGFP)12085 null Rattus norvegicus

Additional Information