Scholarlink

<p>Serine integrases can precisely integrate large DNA constructs into desired chromosomal sites but only if their natural target site is first installed into the recipient genome. Here, to retarget serine integrases to a desired genomic site, we develop a modular integrase (MINT) system for genome editing. Through a combination of structural modeling, single-round directed evolution and screening in human cells, we retargeted the specificity of the serine integrase Bxb1. We demonstrate the therapeutic potential of the MINT system by retargeting Bxb1 to the human <i>AAVS1</i> and <i>TRAC</i> loci, where wild-type Bxb1 has no detectable activity. By combining MINT constructs with both known activity-increasing Bxb1 mutants and zinc-finger DNA-binding domains, we achieve efficiencies of 29% at the <i>AAVS1</i> locus and 35% at the <i>TRAC</i> locus in K562 cells. To further demonstrate clinical potential, we achieved 29% GFP integration efficiencies at the <i>TRAC</i> locus in human T cells.</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

Retargeted serine integrases for one-step, precise integration of large DNA sequences in human cells

  • 期刊论文

  • 发表日期：

    2026年6月29日

查看全文
  • Friedrich Fauser,
  • Sebastian Arangundy-Franklin,
  • Jessica E. Davis,
  • Lifeng Liu,
  • Nicola J. Schmidt,
  • Luis Rodriguez,
  • Danny F. Xia,
  • Nga Nguyen,
  • Yuanyue Zhou,
  • Nicholas A. Scarlott,
  • Lynn N. Truong,
  • Rakshaa Mureli,
  • Irene S. Tan,
  • Satria Sajuthi,
  • Sarah J. Hinkley,
  • Bhakti N. Kadam,
  • Stephen Lam,
  • Bryan Bourgeois,
  • Emily Tait,
  • Mohammad Qasim,
  • Vishvesha Vaidya,
  • Adeline Chen,
  • Andrew Nguyen,
  • Yuri R. Bendaña,
  • David A. Shivak,
  • Patrick Li,
  • Andreas Reik,
  • David E. Paschon,
  • Gregory D. Davis,
  • Jeffrey C. Miller

摘要

Serine integrases can precisely integrate large DNA constructs into desired chromosomal sites but only if their natural target site is first installed into the recipient genome. Here, to retarget serine integrases to a desired genomic site, we develop a modular integrase (MINT) system for genome editing. Through a combination of structural modeling, single-round directed evolution and screening in human cells, we retargeted the specificity of the serine integrase Bxb1. We demonstrate the therapeutic potential of the MINT system by retargeting Bxb1 to the human AAVS1 and TRAC loci, where wild-type Bxb1 has no detectable activity. By combining MINT constructs with both known activity-increasing Bxb1 mutants and zinc-finger DNA-binding domains, we achieve efficiencies of 29% at the AAVS1 locus and 35% at the TRAC locus in K562 cells. To further demonstrate clinical potential, we achieved 29% GFP integration efficiencies at the TRAC locus in human T cells.