A prime editor mouse to model a broad spectrum of somatic mutations in vivo DOI
Zackery A. Ely, Nicolas Mathey‐Andrews,

Santiago Naranjo

et al.

Nature Biotechnology, Journal Year: 2023, Volume and Issue: unknown

Published: May 11, 2023

Language: Английский

Engineered CRISPR prime editors with compact, untethered reverse transcriptases DOI
Julian Grünewald, Bret R. Miller, Regan N. Szalay

et al.

Nature Biotechnology, Journal Year: 2022, Volume and Issue: 41(3), P. 337 - 343

Published: Sept. 26, 2022

Language: Английский

Citations

63

Peptide fusion improves prime editing efficiency DOI Creative Commons
Minja Velimirovic, Larissa C. Zanetti, Max W. Shen

et al.

Nature Communications, Journal Year: 2022, Volume and Issue: 13(1)

Published: June 18, 2022

Abstract Prime editing enables search-and-replace genome but is limited by low efficiency. We present a high-throughput approach, the Peptide Self-Editing sequencing assay (PepSEq), to measure how fusion of 12,000 85-amino acid peptides influences prime show that peptide can enhance editing, prime-enhancing combine productively, and top dual peptide-prime editor increases significantly in multiple cell lines across dozens target sites. Top function increasing translation efficiency serve as broadly useful tools improve

Language: Английский

Citations

48

Adenine Base Editing In Vivo with a Single Adeno-Associated Virus Vector DOI
Han Zhang, Nathan Bamidele, Pengpeng Liu

et al.

GEN Biotechnology, Journal Year: 2022, Volume and Issue: 1(3), P. 285 - 299

Published: June 1, 2022

Base editors (BEs) have opened new avenues for the treatment of genetic diseases. However, advances in delivery approaches are needed to enable disease targeting a broad range tissues and cell types. Adeno-associated virus (AAV) vectors remain one most promising vehicles gene therapies. Currently, BE/guide combinations their promoters exceed packaging limit (∼5 kb) AAVs. Dual-AAV strategies often require high viral doses that impose safety concerns. In this study, we engineered an adenine base editor (ABE) using compact Cas9 from Neisseria meningitidis (Nme2Cas9). Compared with well-characterized Streptococcus pyogenes Cas9-containing ABEs, ABEs Nme2Cas9 (Nme2-ABE) possess distinct protospacer adjacent motif (N4CC) editing window, exhibit fewer off-target effects, can efficiently install therapeutically relevant mutations both human mouse genomes. Importantly, show vivo Nme2-ABE its guide RNA by single AAV vector edit genomic loci revert mutation phenotype adult model tyrosinemia. We anticipate Nme2-ABE, virtue size range, will therapeutic applications improved efficacy due part single-vector system.

Language: Английский

Citations

41

Template-jumping prime editing enables large insertion and exon rewriting in vivo DOI Creative Commons
Chunwei Zheng, Bin Liu, Xiaolong Dong

et al.

Nature Communications, Journal Year: 2023, Volume and Issue: 14(1)

Published: June 8, 2023

Targeted insertion of large DNA fragments holds promise for genome engineering and gene therapy. Prime editing (PE) effectively inserts short (<50 bp) sequences. Employing paired prime guide RNAs (pegRNAs) has enabled PE to better mediate relatively insertions in vitro, but the efficiency larger (>400 remains low vivo application not been demonstrated. Inspired by efficient genomic mechanism retrotransposons, we develop a template-jumping (TJ) approach using single pegRNA. TJ-pegRNA harbors sequence as well two primer binding sites (PBSs), with one PBS matching nicking sgRNA site. TJ-PE precisely 200 bp 500 up 50.5 11.4% efficiency, respectively, enables GFP (~800 expression cells. We transcribe split circular TJ-petRNA vitro via permuted group I catalytic intron non-viral delivery Finally, demonstrate that can rewrite an exon liver tyrosinemia mice reverse disease phenotype. potential insert without double-stranded breaks facilitate mutation hotspot rewriting vivo.

Language: Английский

Citations

35

A prime editor mouse to model a broad spectrum of somatic mutations in vivo DOI
Zackery A. Ely, Nicolas Mathey‐Andrews,

Santiago Naranjo

et al.

Nature Biotechnology, Journal Year: 2023, Volume and Issue: unknown

Published: May 11, 2023

Language: Английский

Citations

30