Targeted RNases: a feasibility study for use in HIV gene therapy.
A targeted RNase would be ideal for gene therapy of several acquired and inherited disorders. Such an RNase may be engineered to contain a ribonucleolytic domain and a specific target RNA binding domain. To demonstrate the feasibility of this approach, an RNase targeted against human immunodeficiency virus (HIV) RNA--Tev-RNase T1--was designed and tested for its use in HIV-1 gene therapy. A human CD4+ T lymphoid (MT4) cell line and human peripheral blood lymphocytes (PBLs) were transduced with retroviral vectors lacking or expressing the tevT1 gene. Expression of enzymatically functional Tev-RNase T1 protein and its lack of toxicity was demonstrated in stable MT4 transductants. Compared with control cells lacking this protein, both transduced MT4 cells and PBLs expressing Tev-RNase T1 delayed HIV-1 replication. Tev-RNase T1 was shown to act after integration, since HIV-1 proviral DNA could be detected, but the amount of HIV-1 RNA produced in MT4 cells and PBLs was significantly decreased. This study demonstrates the feasibility of a targeted RNase strategy for therapeutic use.
Singwi
S
S
Department of Medical Genetics and Microbiology, Faculty of Medicine, University of Toronto, Canada.
Ramezani
A
A
Ding
S F
SF
Joshi
S
S
D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy*; D002460:Cell Line; D002478:Cells, Cultured; D005240:Feasibility Studies; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D012367:RNA, Viral / Q000235:genetics*; D012260:Ribonucleases / Q000235:genetics*; D014162:Transfection / Q000379:methods
D012367:RNA, Viral; D012260:Ribonucleases