Red/ET recombination with chimeric oligonucleotides allows rapid generation of BAC transgenes harboring full-length or truncated huntingtin cDNA

Area of Science:

• Neuroscience
• Genetics
• Molecular Biology

Background:

• Huntington's disease (HD) is a fatal neurodegenerative disorder.
• It is caused by a CAG repeat expansion in the huntingtin (htt) gene.
• Existing HD mouse models do not fully replicate human disease characteristics.

Purpose of the Study:

• To generate novel mouse models of Huntington's disease.
• To overcome challenges in modeling the large and complex htt gene locus.
• To investigate the role of specific neuronal populations in HD pathogenesis.

Main Methods:

• Utilized Red/ET recombination technology.
• Generated four BDNF-BAC transgenes.
• Incorporated full-length or truncated htt cDNA with varying glutamine (Q) repeat lengths (98Q or 15Q).

Main Results:

• Successfully created four distinct BDNF-BAC transgenes harboring human htt cDNA.
• Transgenes express htt with either 98 or 15 glutamine residues.
• BDNF expression directed htt transgene expression to specific neuronal pathways.

Conclusions:

• The generated BDNF-BAC transgenes represent a novel approach to modeling Huntington's disease.
• These models offer a platform to study htt gene expression in targeted neuronal populations.
• Further research using these models can elucidate the contribution of specific cell types to HD.