Human 8-cell embryos enable efficient induction of disease-preventive mutations without off-target effect by cytosine base editor
Protein & Cell
; (12): 416-432, 2023.
Article
en En
| WPRIM
| ID: wpr-982556
Biblioteca responsable:
WPRO
ABSTRACT
Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Palabras clave
Texto completo:
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Índice:
WPRIM
Asunto principal:
Blastocisto
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Citosina
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Apolipoproteína E4
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Sistemas CRISPR-Cas
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Edición Génica
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Heterocigoto
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Mutación
Límite:
Humans
Idioma:
En
Revista:
Protein & Cell
Año:
2023
Tipo del documento:
Article