Open AccessOptimized high‐fidelity 3DPCR to assess potential mitochondrial targeting by activation‐induced cytidine deaminase
Author(s) -
Wu Haiyan,
Zhang Kaili,
Chen Yue,
Li Jinfeng,
Strout Matthew P.,
Gu Xiwen
Publication year - 2020
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12927
Subject(s) - somatic hypermutation , mitochondrial dna , cytidine deaminase , activation induced (cytidine) deaminase , biology , genome , mutagenesis , genetics , mitochondrion , mitochondrial matrix , gene , microbiology and biotechnology , mutation , cytosol , biochemistry , enzyme , antibody , b cell
Here, we develop a high‐fidelity differential DNA denaturation PCR approach and investigate whether activation‐induced cytidine deaminase (AID) can target mitochondrial DNA (mtDNA) in vitro . We demonstrate AID targeting of mtDNA is a rare event. Despite a fraction of AID binding the mitochondrial outer membrane, it is unable to access the mitochondrial matrix or the mtDNA. Thus, the mitochondrial genome is protected from AID‐mediated mutagenesis by physical segregation.