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Published on: June 16, 2017
Generation of Conditional Knockout Alleles for PRUNE-1
Xiaoli Wu1, Louise R Simard1, Hao Ding1,2
1Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, MB R3E 0J9, Canada.
Insights
Researchers created conditional mouse models for PRUNE1, a gene crucial for brain development and function. These models help study PRUNE1
Area of Science:
- Genetics
- Developmental Biology
- Neuroscience
Background:
- PRUNE1, a DHH superfamily protein, is vital for embryogenesis and nervous system development.
- Mutations in PRUNE1 are linked to microcephaly, brain malformations, and neurodegeneration.
- Understanding PRUNE1's cellular function is key for developmental and neurodegenerative disease research.
Purpose of the Study:
- To generate conditional mouse alleles for PRUNE1 to study its function during development and disease.
- To investigate the role of PRUNE1 in Purkinje cell development and survival.
Main Methods:
- Generation of conditional mouse alleles for Prune1 using loxP sites flanking exon 6.
- Crossed conditional alleles with ubiquitous Cre transgenic lines for complete PRUNE1 loss.
- Utilized Purkinje cell-specific Cre line (Pcp2-Cre) to assess PRUNE1's role in Purkinje cells.
- Described an efficient method for generating conditional mouse alleles via zygote electroporation.
Main Results:
- Complete loss of PRUNE1 expression led to embryonic defects mirroring Prune1 null embryos.
- Loss of PRUNE1 in Purkinje cells resulted in Purkinje cell loss, similar to human disease phenotypes.
- The generated conditional alleles are effective tools for studying PRUNE1's spatiotemporal roles.
Conclusions:
- Conditional mouse alleles of Prune1 are valuable tools for developmental and neurodegenerative disease research.
- These models facilitate the study of PRUNE1's role in Purkinje cell development and disease pathogenesis.
- The study provides a method for efficient generation of conditional mouse alleles.
Abstract:
PRUNE1 is a member of the aspartic acid-histidine-histidine (DHH) protein superfamily, which could display an exopolyphosphatase activity and interact with multiple cellular proteins involved in the cytoskeletal rearrangement. It is widely expressed during embryonic development and is essential for embryogenesis. PRUNE1 could also be critical for postnatal development of the nervous system as it was found to be mutated in patients with microcephaly, brain malformations, and neurodegeneration. To determine the cellular function of PRUNE1 during development and in disease, we have generated conditional mouse alleles of the Prune1 in which loxP sites flank exon 6. Crossing these alleles with a ubiquitous Cre transgenic line resulted in a complete loss of PRUNE1 expression and embryonic defects identical to those previously described for Prune1 null embryos. In addition, breeding these alleles with a Purkinje cell-specific Cre line (Pcp2-Cre) resulted in the loss of Purkinje cells similar to that observed in patients carrying a mutation with loss of PRUNE1 function. Therefore, the Prune1 conditional mouse alleles generated in this study provide important genetic tools not only for dissecting the spatial and temporal roles of PRUNE1 during development but also for understanding the pathogenic role of PRUNE1 dysfunction in neurodegenerative or neurodevelopmental disease. In addition, from this work, we have described an approach that allows one to efficiently generate conditional mouse alleles based on mouse zygote electroporation.
