Inhibition of ampicillin-resistant bacteria by novel mono-DNAzymes and di-DNAzyme targeted to beta-lactamase mRNA.
In view of the weakness of antibiotics and the properties of antisense drugs, we applied DNAzymes to the field of drug resistance in bacteria. Two 10-23 mono-DNAzymes (Dz1, Dz2) and a di-DNAzyme (Dz1-2) targeted to beta-lactamase mRNA were designed to determine to what degree the growth of ampicillin-resistant bacteria (TEM-1, TEM-3) was inhibited. All three DNAzymes can play a role both in vitro and in vivo. In vitro, they exhibited high catalytic efficiency (kcat/KM) of 63.5, 91.1, and 30.8 pM(-1) min(-1), respectively, under multiple-turnover conditions. In vivo, after 9 hours' incubation, the degree of inhibition of Dz1, Dz2, and Dz1-2 for TEM-1 bacteria was 27.2%, 39.6%, and 57.7%, respectively, and that for TEM-3 bacteria was 39.1%, 44%, and 62.6%, respectively. Dz1-2 showed the greatest inhibiting effect, demonstrating in vivo activity may be increased by constructing multiple-target DNAzymes. The results indicated a potential possibility for DNAzymes to act as a new type of antibacterial or a tool of gene functional analysis for prokaryocytes.
Key Lab for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, Changchun City, Jilin Province, 130023, P. R. China.
Fei
F
Chen
Zhe
Z
Li
Ruijian
R
Wang
Bin
B
Liu
Zhen
Z
Zeng
Hongying
H
Zhang
Jin
J
Zhang
D000668:Ampicillin Resistance / Q000235:genetics*; D001419:Bacteria / Q000187:drug effects* / Q000235:genetics*; D001483:Base Sequence; D021881:DNA, Catalytic / Q000096:biosynthesis / Q000494:pharmacology*; D016503:Drug Delivery Systems / Q000379:methods*; D015195:Drug Design; D018274:Electroporation; D007700:Kinetics; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics*
D021881:DNA, Catalytic; D012333:RNA, Messenger; D001618:beta-Lactamases