Scholarlink

<p>CRISPR–Cas3 represents a mechanistically distinct genome-editing system compared to Cas9 that generates long-range deletions rather than small indels, thereby reducing the risk of residual protein function from in-frame mutations. Here we evaluated CRISPR–Cas3 to correct mutations in the <i>TTR</i> gene causing transthyretin amyloidosis, a systemic proteinopathy where loss of mutant TTR in the liver offers therapeutic benefit. Through CRISPR RNA optimization we achieved 58.9% ± 0.5% editing at the <i>TTR</i> locus in vitro, inducing large deletions that abolished TTR expression. Cas3 generated mostly directional deletions up to 75 kb without reproducible off-target mutations, in contrast to Cas9, which induced indels at several off-target sites. In vivo, a single lipid-nanoparticle-based treatment achieved 48.7% ± 1.1% hepatic editing and reduced serum TTR levels by 80.1% ± 4.6%. Deletion size was limited to 21 kb. In <i>TTR</i> exon-humanized mice, Cas3 editing reduced serum TTR without in-frame mutations and attenuated macrophage-associated TTR deposition. These findings highlight Cas3 as an efficient and distinct sytem for in vivo genome editing.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

CRISPR–Cas3-based editing for targeted deletions in a mouse model of transthyretin amyloidosis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年1月5日

查看全文
  • Saeko Ishida,
  • Yusuke Sato,
  • Keisuke Chosa,
  • Eri Ezawa,
  • Yuko Yamauchi,
  • Masaaki Oyama,
  • Hiroko Kozuka-Hata,
  • Rina Ito,
  • Rikako Sato,
  • Masatoshi Maeki,
  • Tomo-o Ishikawa,
  • Kenichi Yamamura,
  • Kohei Takeshita,
  • Kensuke Yamaguchi,
  • Yuta Kochi,
  • Fumitaka Hashiya,
  • Yiwei Liu,
  • Naoko Abe,
  • Hiroshi Abe,
  • Yoshiki Sekijima,
  • Kazuto Yoshimi,
  • Tomoji Mashimo

摘要

CRISPR–Cas3 represents a mechanistically distinct genome-editing system compared to Cas9 that generates long-range deletions rather than small indels, thereby reducing the risk of residual protein function from in-frame mutations. Here we evaluated CRISPR–Cas3 to correct mutations in the TTR gene causing transthyretin amyloidosis, a systemic proteinopathy where loss of mutant TTR in the liver offers therapeutic benefit. Through CRISPR RNA optimization we achieved 58.9% ± 0.5% editing at the TTR locus in vitro, inducing large deletions that abolished TTR expression. Cas3 generated mostly directional deletions up to 75 kb without reproducible off-target mutations, in contrast to Cas9, which induced indels at several off-target sites. In vivo, a single lipid-nanoparticle-based treatment achieved 48.7% ± 1.1% hepatic editing and reduced serum TTR levels by 80.1% ± 4.6%. Deletion size was limited to 21 kb. In TTR exon-humanized mice, Cas3 editing reduced serum TTR without in-frame mutations and attenuated macrophage-associated TTR deposition. These findings highlight Cas3 as an efficient and distinct sytem for in vivo genome editing.