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Background <p>HD is a devastating neurodegenerative disorder caused by the expansion of CAG repeats in the <i>HTT</i>. Silencing the expression of mutated proteins is a therapeutic direction to rescue HD patients, and recent advances in gene editing technology such as CRISPR/CasRx have opened up new avenues for therapeutic intervention.</p> Methods <p>The CRISPR/CasRx system was employed to target human HTT exon 1, resulting in an efficient knockdown of <i>HTT</i> mRNA. This therapeutic effect was substantiated in various models: HEK 293&#xa0;T cell, the HD 140Q-KI mouse, and the HD-KI pig model. The efficiency of the knockdown was analyzed through Western blot and RT-qPCR. Additionally, neuropathological changes were examined using Western blot, immunostaining, and RNA sequencing. The impact on motor abilities was assessed via behavioral experiments, providing a comprehensive evaluation of the treatment's effectiveness.</p> Results <p>CRISPR/CasRx system can significantly reduce <i>HTT</i> mRNA levels across various models, including HEK 293&#xa0;T cells, HD 140Q-KI mice at various disease stages, and HD-KI pigs, and resulted in decreased expression of mHTT. Utilizing the CRISPR/CasRx system to knock down <i>HTT</i> RNA has shown to ameliorate gliosis in HD 140Q-KI mice and delay neurodegeneration in HD pigs.</p> Conclusions <p>These findings highlight the effectiveness of the RNA-targeting CRISPR/CasRx as a potential therapeutic strategy for HD. Furthermore, the success of this approach provides valuable insights and novel avenues for the treatment of other genetic disorders caused by gene mutations.</p>

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RNA-Targeting CRISPR/CasRx system relieves disease symptoms in Huntington’s disease models

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年1月13日

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  • Yingqi Lin,
  • Caijuan Li,
  • Yizhi Chen,
  • Jiale Gao,
  • Jiawei Li,
  • Chunhui Huang,
  • Zhaoming Liu,
  • Wei Wang,
  • Xiao Zheng,
  • Xichen Song,
  • Jianhao Wu,
  • Jiaxi Wu,
  • Oscar Junhong Luo,
  • Zhuchi Tu,
  • Shihua Li,
  • Xiao-Jiang Li,
  • Liangxue Lai,
  • Sen Yan

摘要

Background

HD is a devastating neurodegenerative disorder caused by the expansion of CAG repeats in the HTT. Silencing the expression of mutated proteins is a therapeutic direction to rescue HD patients, and recent advances in gene editing technology such as CRISPR/CasRx have opened up new avenues for therapeutic intervention.

Methods

The CRISPR/CasRx system was employed to target human HTT exon 1, resulting in an efficient knockdown of HTT mRNA. This therapeutic effect was substantiated in various models: HEK 293 T cell, the HD 140Q-KI mouse, and the HD-KI pig model. The efficiency of the knockdown was analyzed through Western blot and RT-qPCR. Additionally, neuropathological changes were examined using Western blot, immunostaining, and RNA sequencing. The impact on motor abilities was assessed via behavioral experiments, providing a comprehensive evaluation of the treatment's effectiveness.

Results

CRISPR/CasRx system can significantly reduce HTT mRNA levels across various models, including HEK 293 T cells, HD 140Q-KI mice at various disease stages, and HD-KI pigs, and resulted in decreased expression of mHTT. Utilizing the CRISPR/CasRx system to knock down HTT RNA has shown to ameliorate gliosis in HD 140Q-KI mice and delay neurodegeneration in HD pigs.

Conclusions

These findings highlight the effectiveness of the RNA-targeting CRISPR/CasRx as a potential therapeutic strategy for HD. Furthermore, the success of this approach provides valuable insights and novel avenues for the treatment of other genetic disorders caused by gene mutations.