Phi29 pRNA vector for efficient escort of hammerhead ribozyme targeting survivin in multiple cancer cells.
Ribozymes are potential therapeutic agents which suppress specific genes in disease-affected cells. Ribozymes have high substrate cleavage efficiency, yet their medical application has been hindered by RNA degradation, aberrant cell trafficking, or misfolding when fused to a carrier. In this study, we constructed a chimeric ribozyme escorted by the motor pRNA of bacteriophage phi29 to achieve proper folding and enhanced stability. A pRNA molecule contains an interlocking loop domain and a 5'/3' helical domain, which fold independently of one another. When a ribozyme is connected to the helical domain, the chimeric pRNA/ribozyme reorganizes into a circularly permuted form, and the 5'/3' ends are relocated and buried in the original 71'/75' positions. Effective silencing of the anti-apoptotic gene survivin by an appropriately designed chimeric ribozyme, as demonstrated at mRNA and protein levels, led to programmed cell death in various human cancer cell lines, including breast, prostate, cervical, nasopharyngeal, and lung, without causing significant non-specific cytotoxicity. Through the interlocking interaction of right and left loops, monomer pRNA/ribozyme chimeras can be incorporated into multi-functional dimer, trimer and hexamer complexes for specific gene delivery. Using the phi29 motor pRNA as an escort may revive the ribozyme's strength in medical application.
Liu
Hongyun
H
Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Guo
Songchuan
S
Roll
Richard
R
Li
Jie
J
Diao
Zhijuan
Z
Shao
Ningsheng
N
Riley
Mark R
MR
Cole
Alexander M
AM
Robinson
J Paul
JP
Snead
Nicholas M
NM
Shen
Guanxin
G
Guo
Peixuan
P
D017209:Apoptosis; D017103:Bacillus Phages / Q000235:genetics*; D015153:Blotting, Western; D002465:Cell Movement; D019281:Dimerization; D016503:Drug Delivery Systems; D005434:Flow Cytometry; D015870:Gene Expression; D020868:Gene Silencing; D018390:Gene Targeting; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D009336:Necrosis; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D012317:RNA Phages / Q000235:genetics*; D016337:RNA, Catalytic / Q000235:genetics / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D000077022:Survivin; D014162:Transfection; D014407:Tumor Cells, Cultured
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016337:RNA, Catalytic; D012333:RNA, Messenger; D000077022:Survivin; C427204:hammerhead ribozyme