Adenine base editing and prime editing of chemically derived hepatic progenitors rescue genetic liver disease

Kim, Yo-Han; Hong, Sung-Ah; Yu, Jong Pil; Eom, Jeongyun; Jang, Kiseok; Yoon, Sangtae; Hong, Da Hee; Seo, Daekwan; Lee, Seu-Na; Woo, Jae Sung; Jeong, Jae‐Weon; Bae, Sangsu; Choi, Dongho

doi:10.1016/j.stem.2021.04.010

Cited by 46 publications

(36 citation statements)

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“…Consistent with previous studies, we revealed here with WGS analysis that long term overexpression of PE2 produced negligible off-target effects in hESCs. These results strongly support further applications of PEs for therapeutic editing in clinical studies ( 41 , 43 , 44 ).…”

Section: Discussionsupporting

confidence: 80%

Comprehensive analysis of prime editing outcomes in human embryonic stem cells

Habib

Hwang

et al. 2022

Nucleic Acids Research

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Prime editing is a versatile and precise genome editing technique that can directly copy desired genetic modifications into target DNA sites without the need for donor DNA. This technique holds great promise for the analysis of gene function, disease modeling, and the correction of pathogenic mutations in clinically relevant cells such as human pluripotent stem cells (hPSCs). Here, we comprehensively tested prime editing in hPSCs by generating a doxycycline-inducible prime editing platform. Prime editing successfully induced all types of nucleotide substitutions and small insertions and deletions, similar to observations in other human cell types. Moreover, we compared prime editing and base editing for correcting a disease-related mutation in induced pluripotent stem cells derived form a patient with α 1-antitrypsin (A1AT) deficiency. Finally, whole-genome sequencing showed that, unlike the cytidine deaminase domain of cytosine base editors, the reverse transcriptase domain of a prime editor does not lead to guide RNA-independent off-target mutations in the genome. Our results demonstrate that prime editing in hPSCs has great potential for complementing previously developed CRISPR genome editing tools.

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Section: Discussionsupporting

confidence: 80%

Comprehensive analysis of prime editing outcomes in human embryonic stem cells

Habib

Hwang

et al. 2022

Nucleic Acids Research

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“…These unique properties of prime editing are based on the delivery of a prime editor (PE), consisting of a Cas9-reverse transcriptase fusion protein (hereafter referred to as PE2), along with the prime editing guide RNA (pegRNA) that specifies both the genomic target as well as the desired edit to be written directly into the genome. Prime editing has tremendous potential for treatment of disease-causing mutations, as well as generation of disease models, both in vitro and in vivo ( Schene et al, 2020 ; Jang et al, 2021 ; Kim et al, 2021 ; Liu et al, 2021 ; Park et al, 2021 ; Petri et al, 2021 ; Qian et al, 2021 ). However, the use of prime editing is currently challenged by low efficacy, leading to time-consuming optimization and/or screening approaches in order to achieve satisfactory editing activities ( Liu et al, 2020 ; Schene et al, 2020 ; Chemello et al, 2021 ; Kim et al, 2021 ; Petri et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

“…Prime editing has tremendous potential for treatment of disease-causing mutations, as well as generation of disease models, both in vitro and in vivo ( Schene et al, 2020 ; Jang et al, 2021 ; Kim et al, 2021 ; Liu et al, 2021 ; Park et al, 2021 ; Petri et al, 2021 ; Qian et al, 2021 ). However, the use of prime editing is currently challenged by low efficacy, leading to time-consuming optimization and/or screening approaches in order to achieve satisfactory editing activities ( Liu et al, 2020 ; Schene et al, 2020 ; Chemello et al, 2021 ; Kim et al, 2021 ; Petri et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

piggyPrime: High-Efficacy Prime Editing in Human Cells Using piggyBac-Based DNA Transposition

et al. 2021

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Prime editing is a novel genome editing technology that allows a wide range of tailored genomic alterations. Prime editing does not involve homologous recombination, but suffers from low efficacy. Here, we demonstrate piggyPrime, a transfected single-vector system based on piggyBac DNA transposition for genomic integration of all prime editing components in human cells allowing easy and effective transgenesis with prime editing efficacies up to 100% in cell lines.

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“…In terms of clinical research, a PE can repair genetic mutations in sickle cell disease and Tay-Sachs disease ( Anzalone et al, 2019 ). PEs can also achieve base replacement to treat tyrosinemia in mice ( Kim et al, 2021a ).…”

Section: Rna-guided Dna-targeting Effectorsmentioning

confidence: 99%

Development and Application of CRISPR-Cas Based Tools

2022

Front. Cell Dev. Biol.

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Abundant CRISPR-Cas systems in nature provide us with unlimited valuable resources to develop a variety of versatile tools, which are powerful weapons in biological discovery and disease treatment. Here, we systematically review the development of CRISPR-Cas based tools from DNA nuclease to RNA nuclease, from nuclease dependent-tools to nucleic acid recognition dependent-tools. Also, considering the limitations and challenges of current CRISPR-Cas based tools, we discuss the potential directions for development of novel CRISPR toolkits in the future.

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Adenine base editing and prime editing of chemically derived hepatic progenitors rescue genetic liver disease

Cited by 46 publications

References 50 publications

Comprehensive analysis of prime editing outcomes in human embryonic stem cells

Comprehensive analysis of prime editing outcomes in human embryonic stem cells

piggyPrime: High-Efficacy Prime Editing in Human Cells Using piggyBac-Based DNA Transposition

Development and Application of CRISPR-Cas Based Tools

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