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Chromatin context-dependent regulation and epigenetic manipulation of prime editing.
Li, Xiaoyi; Chen, Wei; Martin, Beth K; Calderon, Diego; Lee, Choli; Choi, Junhong; Chardon, Florence M; McDiarmid, Troy A; Daza, Riza M; Kim, Haedong; Lalanne, Jean-Benoît; Nathans, Jenny F; Lee, David S; Shendure, Jay.
Afiliação
  • Li X; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA. Electronic address: xyli10@uw.edu.
  • Chen W; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Molecular Engineering and Sciences Institute, University of Washington, Seattle, WA 98195, USA.
  • Martin BK; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Calderon D; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Lee C; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Choi J; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Howard Hughes Medical Institute, Seattle, WA 98195, USA.
  • Chardon FM; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • McDiarmid TA; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Daza RM; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Kim H; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Lalanne JB; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Nathans JF; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Medical Scientist Training Program, University of Washington, Seattle, WA 98195, USA.
  • Lee DS; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
  • Shendure J; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Howard Hughes Medical Institute, Seattle, WA 98195, USA; Brotman Baty Institute for Precision Medicine, Seattle, WA 98195, USA; Allen Discovery Center for Cell Lineage Tracing, Seattle, WA 98109, USA; Seattle Hub for Sy
Cell ; 187(10): 2411-2427.e25, 2024 May 09.
Article em En | MEDLINE | ID: mdl-38608704
ABSTRACT
We set out to exhaustively characterize the impact of the cis-chromatin environment on prime editing, a precise genome engineering tool. Using a highly sensitive method for mapping the genomic locations of randomly integrated reporters, we discover massive position effects, exemplified by editing efficiencies ranging from ∼0% to 94% for an identical target site and edit. Position effects on prime editing efficiency are well predicted by chromatin marks, e.g., positively by H3K79me2 and negatively by H3K9me3. Next, we developed a multiplex perturbational framework to assess the interaction of trans-acting factors with the cis-chromatin environment on editing outcomes. Applying this framework to DNA repair factors, we identify HLTF as a context-dependent repressor of prime editing. Finally, several lines of evidence suggest that active transcriptional elongation enhances prime editing. Consistent with this, we show we can robustly decrease or increase the efficiency of prime editing by preceding it with CRISPR-mediated silencing or activation, respectively.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cromatina / Epigênese Genética / Sistemas CRISPR-Cas / Edição de Genes Limite: Humans Idioma: En Revista: Cell Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Cromatina / Epigênese Genética / Sistemas CRISPR-Cas / Edição de Genes Limite: Humans Idioma: En Revista: Cell Ano de publicação: 2024 Tipo de documento: Article