[Correction of the pathogenic mutation in the G6PC3 gene by adenine base editing in mutant embryos].
Zhonghua Xue Ye Xue Za Zhi
; 44(4): 308-315, 2023 Apr 14.
Article
en Zh
| MEDLINE
| ID: mdl-37357000
ABSTRACT
Objective:
To determine whether the adenine base editor (ABE7.10) can be used to fix harmful mutations in the human G6PC3 gene.Methods:
To investigate the safety of base-edited embryos, off-target analysis by deep sequencing was used to examine the feasibility and editing efficiency of various sgRNA expression vectors. The human HEK293T mutation models and human embryos were also used to test the feasibility and editing efficiency of correction.Results:
â The G6PC3(C295T) mutant cell model was successfully created. â¡In the G6PC3(C295T) mutant cell model, three distinct Re-sgRNAs were created and corrected, with base correction efficiency ranging from 8.79% to 19.56% . ⢠ABE7.10 could successfully fix mutant bases in the human pathogenic embryo test; however, base editing events had also happened in other locations. ⣠With the exception of one noncoding site, which had a high safety rate, deep sequencing analysis revealed that the detection of 32 probable off-target sites was <0.5% .Conclusion:
This study proposes a new base correction strategy based on human pathogenic embryos; however, it also produces a certain nontarget site editing, which needs to be further analyzed on the PAM site or editor window.Palabras clave
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Sistemas CRISPR-Cas
/
Edición Génica
Idioma:
Zh
Revista:
Zhonghua Xue Ye Xue Za Zhi
Año:
2023
Tipo del documento:
Article