129/Ola mice carrying a null mutation in PrP that abolishes mRNA production are developmentally normal

Manson, Jean; Clarke, Alan R.; Hooper, Martin; Aitchison, L; McConnell, I.; Hope, James

doi:10.1007/bf02780662

Cited by 547 publications

(436 citation statements)

References 35 publications

Supporting

Mentioning

420

Contrasting

Unclassified

Order By: Relevance

“…The suggestion that there may be a synaptic deficiency in Prnp °/° mice [42,43] has not been confirmed [44]. The claim that aged mice (with a mixed genetic background) develop ataxia and suffer a loss of Purkinje cells [45] as a consequence of PrP gene disruption is not consistent with previous investigations on independently generated Prnp °/° mouse lines [41,46]. Because the phenotype might be due to the undefined, mixed genetic background of the knockout mice [47], it is necessary to show that complementation with a PrP transgene restores the normal phenotype.…”

Section: Resistance To Scrapie Of Mice Devoid Of Prp Cmentioning

confidence: 87%

Molecular biology of transmissible spongiform encephalopathies

1996

View full text Add to dashboard Cite

The prion, the transmissible agent that causes spongiform encephalopathies such as serapie, bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease, is believed to be devoid of nucleic acid and identical with PrP so, a modified form of the normal host protein PrP ¢ which is encoded by the single cop~v gene Prnp. The 'protein only" hypothesis proposes that PrP ~c, when introduced into a normal host, causes the conversion of PrP c into prpS~; it therefore predicts that an animal devoid of PrP c should be resistant to prion diseases. We generated homozygous Prnp °1° ('PrP knockout') mice and showed that, after inoculation with prions, they remained free of scrapie for at least 2 years while wild-type controls all died within 6 months. There was no propagation of prions in the Prnp °t° animals. Surprisingly, heterozygous Prnp °t+ mice, which express PrP c at about half the normal level, also showed enhanced resistance to scrapie disease despite high levels of infectious agent and PrP ~c in the brain early on. After introduction of murine PrP transgenes Prnp °t° mice became highly susceptible to mouse but not to hamster prions, while the insertion of Syrian hamster PrP transgenes rendered them susceptible to hamster but to a much lesser extent to mouse prions. These complementation experiments paved the way to the application of reverse genetics. We have prepared animals transgenic for genes encoding PrP with amino terminal deletions of various lengths and have found that PrP lacking 48 amino proximal amino acids, which comprise four of the five octa repeats of PrP, is still biologically active.

show abstract

Section: Resistance To Scrapie Of Mice Devoid Of Prp Cmentioning

confidence: 87%

Molecular biology of transmissible spongiform encephalopathies

1996

View full text Add to dashboard Cite

show abstract

“…The generation of two lines of mice by disrupting Prnp expression was carried out in the early 1990s: Zürich I (outbred) (Bueler et al, 1992) and Edinburgh I (inbred) (Manson et al, 1994) lines (see (Weissmann and Aguzzi, 1999) for review). Prnpknockout mice are resistant to prion infection (Bueler et al, 1993) but surprisingly do not show major phenotypical defects.…”

Section: Prnp-deficient Micementioning

confidence: 99%

New insights into cellular prion protein (PrPc) functions: The “ying and yang” of a relevant protein

Nicolas

Gavı́n

Rı́o

2009

Brain Research Reviews

View full text Add to dashboard Cite

show abstract

“…Separate cultures were set up in triplicate microtiter wells or 48-well plate for proliferation and cytokine assays. For proliferation assays, cultures were pulsed at 48 h and 72 h with 0.5 µCi of 3 …”

Section: In Vitro T-cell Responses To Mitogen Stimulationmentioning

confidence: 99%

“…Disruption of PrP C expression in mice, a species that does not naturally contract prion diseases, results in no apparent developmental abnormalities [2][3][4][5] . However, the impact of ablating PrP C function in natural host species of prion diseases is unknown.…”

Section: Introductionmentioning

confidence: 99%

Production of cattle lacking prion protein

et al. 2006

View full text Add to dashboard Cite

Prion diseases are caused by propagation of misfolded forms of the normal cellular prion protein PrP C , such as PrP BSE in bovine spongiform encephalopathy (BSE) in cattle and PrP CJD in Creutzfeldt-Jakob disease (CJD) in humans 1 . Disruption of PrP C expression in mice, a species that does not naturally contract prion diseases, results in no apparent developmental abnormalities [2][3][4][5] . However, the impact of ablating PrP C function in natural host species of prion diseases is unknown. Here we report the generation and characterization of PrP C -deficient cattle produced by a sequential gene-targeting system 6 . At over 20 months of age, the cattle are clinically, physiologically, histopathologically, immunologically and reproductively normal. Brain tissue homogenates are resistant to prion propagation in vitro as assessed by protein misfolding cyclic amplification 7 . PrP C -deficient cattle may be a useful model for prion research and could provide industrial bovine products free of prion proteins.To generate PrP C -deficient (PRNP −/− ) cattle, we transfected a male Holstein primary fetal fibroblast line 6594 with first and second knockout (KO) vectors (pBPrP(H)KOneo and pBPrP (H)KOpuro vectors) 6 to sequentially disrupt the two alleles of PRNP. PRNP −/− fetal cell lines were established at 40-60 d of gestation and three of the PRNP −/− fetal cell lines (5211, 5232 and 4296) were recloned to produce calves (Table 1 and Fig. 1a). To verify that the calves possess the PRNP −/− genotype, we collected ear biopsies and established fibroblast cell lines for genotyping. Genotyping was done by genomic PCR specific to each gene targeting event 6 (primer pairs: neoF7 × neoR7 and puroF14 × puroR14, Fig. 1b COMPETING INTERESTS STATEMENTThe authors declare competing financial interests (see the Nature Biotechnology website for details).Reprints and permissions information is available online at http://npg.nature.com/reprintsandpermissions/ NIH Public Access Fig. 1c) and confirmed the disruption of PRNP-specific mRNA expression in PRNP −/− calves. For protein expression analysis, we performed PrP-specific western blot analyses on fibroblasts (Fig. 1d), peripheral blood lymphocytes (Fig. 1e) and brain stem (Fig. 1f) from wild-type and PRNP −/− calves using the mouse anti-bovine PrP monoclonal antibody F89. We detected PrP-specific bands in the wild-type calves, whereas no reaction was observed in PRNP −/− calves and negative control mouse fibroblasts. These data clearly demonstrate that the PRNP gene is functionally inactivated in the PRNP −/− calves.PRNP −/− cattle were monitored for growth and general health status from birth to 20 months of age. Mean birth weight was 46 kg and average daily gain was 0.91 kg/d to 10 months. Both values were in the normal range for Holstein bulls. Serum chemistry was evaluated at 6 months of age and compared with published reference ranges. All the values for PRNP −/− calves (n = 12) were well within the reference range (Supplementary Table 1) and obvious abnormalities w...

show abstract

129/Ola mice carrying a null mutation in PrP that abolishes mRNA production are developmentally normal

Cited by 547 publications

References 35 publications

Molecular biology of transmissible spongiform encephalopathies

Molecular biology of transmissible spongiform encephalopathies

New insights into cellular prion protein (PrPc) functions: The “ying and yang” of a relevant protein

Production of cattle lacking prion protein

Contact Info

Product

Resources

About