ABSTRACT
Molecular profiling of normal tissues is a regular and necessary step when developing systems for expression analyses in biological samples, including diagnostic panels for various diseases and conditions. Yet there are still no rigorous criteria to allow precise typing of normal tissues. A main problem is that the methods employed in diagnostic expression testing are difficult to standardize. While various technologies, instruments, and reagents are available, universal protocols of handling biological material are lacking, thus impairing the reproducibility of data from independent studies. The review describes a new approach to standardizing circulating microRNA studies in forensic biology, which has relatively recently (7-8 years ago) come to employ RNA markers in molecular typing of tissues and biological fluids. Forensic biology is now one of the few disciplines where several panels of tissue mRNA markers have been developed within a short period of time and a number of specific microRNA markers have been established and validated for several biological fluids. To allow their successful use, new protocols have been combined with the available, rigidly standardized system of genetic personal identification. Although a ready diagnostic product has still not been obtained with this well-working approach, the apparent efficiency of the standardization methods clearly demonstrates that the problem is possible to solve in other biomedical fields, including those where RNA-based diagnostic protocols are still under development.
Subject(s)
Circulating MicroRNA/blood , Forensic Sciences/methods , Polymerase Chain Reaction/standards , RNA, Messenger/blood , Biomarkers/metabolism , Circulating MicroRNA/genetics , Female , Forensic Sciences/instrumentation , Humans , Male , Nipple Discharge/chemistry , Nipple Discharge/metabolism , Polymerase Chain Reaction/instrumentation , Polymerase Chain Reaction/methods , RNA, Messenger/genetics , Reproducibility of Results , Saliva/chemistry , Saliva/metabolism , Semen/chemistry , Semen/metabolism , Sensitivity and Specificity , Vaginal Discharge/diagnosis , Vaginal Discharge/genetics , Vaginal Discharge/metabolismABSTRACT
A new target has been revealed for therapy for herpes simplex type 2. The target is RS1 mRNA whose activation is a key stage in regulating the expression of the early genes of herpes simplex virus type 2 (HSV-2). The targeted knockdown of the function of this gene by small interfering RNAs (siRNAs) has been found to result in the complete suppression of HSV-2 infection. The designed interfering RNAs are able to interact with both HSV-1 and HSV-2. The findings suggest that siRNAs have protective properties against the cytopathic effect of HSV-2 and cause no toxicity to infected Vero cells.
Subject(s)
Herpesvirus 1, Human/drug effects , Herpesvirus 2, Human/genetics , Immediate-Early Proteins/genetics , RNA, Small Interfering/genetics , Animals , Antiviral Agents/administration & dosage , Chlorocebus aethiops , Gene Expression Regulation, Viral , Gene Knockdown Techniques , Herpesvirus 2, Human/drug effects , Humans , RNA, Messenger/antagonists & inhibitors , RNA, Messenger/genetics , RNA, Small Interfering/administration & dosage , Vero Cells , Virus Replication/drug effectsABSTRACT
The use of the RNA interference technique yielded data on the antiviral activity of small interfering RNA (siRNA) oligonucleotides against hepatitis C virus (HCV) infection in the pig embryo kidney (SPEV) cell cultures. The RNA interference technique is based on the specific recognition of the mRNA target by using the specially designed siRNA (19-22 bp) oligonucleotides. In particular, it was shown that siRNA added to the monolayer of HCV-infected SPEV cells resulted in the protection of the infected cells against the cytopathogenic activity of the virus. The results were confirmed in the experiments that demonstrated the ability of RNA oligonucleotides to reduce the production of infectious (cytopathogenic) HCV by infected SPEV cells in early-stage infection.
Subject(s)
Antiviral Agents/pharmacology , Hepacivirus/drug effects , RNA, Small Interfering/pharmacology , Virus Replication/drug effects , Animals , Cell Line , Hepatitis C/therapy , Humans , SwineABSTRACT
Peripheral blood mononuclear cells of 24-70% individuals infected with HTLV-1 contain defective proviruses (dp) in addition to the full size ones. Most of them remain silent lacking regions sufficient for viral genes transcription except those activated under cell promoter or retaining viral open reading frames (orfs). It is still unclear whether these proviruses are associated with the development of T-cell leukemia in adults, tropic spastic paresis, or myelopathy. Classification of previously reported dp is presented, their origin and possible function in human HTLV-1 associated diseases are discussed.