Degradation and stable maintenance of adeno-associated virus inverted terminal repeats in E. coli

被引：0

作者：

Radukic, Marco T. ^{[1
]}

Le, Dinh To ^{[1
,3
]}

Krassuski, Timo ^{[1
]}

Borchert, Philipp ^{[1
]}

Leach, David R. F. ^{[2
]}

Mueller, Kristian M. ^{[1
]}

机构：

[1] Bielefeld Univ, Fac Technol, Cellular & Mol Biotechnol, Univ Str 25, D-33615 Bielefeld, North Rhine Wes, Germany

[2] Univ Edinburgh, Inst Cell Biol, Sch Biol Sci, Kings Bldg,Mayfield Rd, Edinburgh EH9 3BF, Scotland

[3] Univ Med Ctr Hamburg Eppendorf, Dept Med, D-20246 Hamburg, Germany

来源：

NUCLEIC ACIDS RESEARCH | 2024年 / 53卷 / 02期

关键词：

ESCHERICHIA-COLI; RECOMBINANT PLASMID; DNA HAIRPINS; PALINDROME; REPLICATION; GENE; MECHANISM; DELETION; RESCUE;

D O I：

10.1093/nar/gkae1170

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Current plasmid propagation in E. coli compromises large inverted repeats, such as inverted terminal repeats (ITRs) of adeno-associated virus (AAV). Direct long-read sequencing analyses upon varying strains and culture conditions revealed ITR instability caused by a slipped misalignment mechanism, although other mechanism probably contribute. ITRs stabilized in absence of SbcC, which is part of the SbcCD nuclease complex, a human Mre11-Rad50 homolog, or at elevated growth temperatures (e.g. 42 degrees C), with a combination being optimal. Resulting full ITR transgene plasmids improved rAAV yield and purity in HEK-293 productions. The findings advance plasmid biology, cloneable sequences and therapeutic AAV manufacturing.

引用

页数：6