The Center for RNA Interference and Non-Coding RNAs (RNA Center), established under the Red and Charline McCombs Institute for the Early Detection and Treatment of Cancer, is a unique collaborative initiative among MD Anderson Cancer Center, Baylor College of Medicine, The University of Texas Health Science Center at Houston, Rice University and the University of Houston that will focus on gaining insights into the roles of newly discovered RNAs in cancer initiation, progression and dissemination.
Our goal is to identify, engineer and accelerate breakthroughs in non-coding RNA (ncRNA) discoveries leading to cancer biomarkers and therapeutics.
Our short-term objective is to build on existing clinical research at MD Anderson Cancer Center through collaborations and membership to the RNA Center to create a ncRNA-centric effort to drive discovery of molecular markers of cancer by evaluating, co-developing, facilitating and disseminating novel ncRNA technologies.
Our long-term objective is to create an institution-wide ncRNA resource to collaborate and lend expertise in the ncRNA and RNAi areas of clinical and basic research.
Rahul Mitra, Ph.D.
Program Director, RNA Center
Email: rmitra@mdanderson.org or rnacenter@mdanderson.org
The highly efficient delivery of short-interfering RNA (siRNA, shown in red) is illustrated.. The siRNA was packaged in neutral nanoliposomes which were intravenously injected into mouse models. The target: ovarian cancer cells (blue).
Overview
With promise in broad areas ranging from relief of cancer-related chronic pain to management of deadly brain metastasis, RNAi has potential applications for every type of cancer. RNAi offers a powerful and highly specific method for shutting off genes that promote cancer growth. Our researchers aim to develop this potential across the full continuum of cancer care.
MicroRNA and other short or long non-coding RNAs alterations are involved in the initiation, progression and metastases of human cancer. The main molecular alterations are represented by variations in gene expression, usually mild and with consequences for a vast number of target protein coding genes. The causes of the widespread differential expression of non-coding RNAs in malignant cells, as compared to normal cells, can be explained by the location of these genes in cancer-associated genomic regions, by epigenetic mechanisms and by alterations in the processing machinery.
Expression profiling of microRNA and other short or long non-coding RNAs in human tumors has identified signatures associated with diagnosis, staging, progression, prognosis and response to treatment. In addition, profiling has been exploited to identify non-coding RNAs that may represent downstream targets of activated oncogenic pathways or are targeting protein coding genes involved in cancer. Recent studies have demonstrated that miRNAs and non-coding ultraconserved genes are main candidates for the elusive class of cancer predisposing genes and that other types of non-coding RNAs participate in the genetic puzzle giving rise to the malignant phenotype. These discoveries could be exploited for the development of useful markers for diagnosis and prognosis, as well as for the development of new RNA-based cancer therapies.
RNAi at a Glance
RNAi is a type of gene therapy that “silences” genes by disrupting the protein-making process. That process is essentially explained in a statement which scientists have dubbed the “central dogma of molecular biology”: DNA makes RNA makes proteins.
DNA lives in the cell nucleus and consists of thousands of genes, each one holding the code, or “blueprint,” for building a particular protein molecule and thus ensuring a certain cell function performed by that protein. (Proteins carry out virtually every cell function, including the activation of other genes.) When a specific protein is needed by the body, its genetic code is copied from DNA; the “copy” is a single molecular strand known as messenger RNA (mRNA). This process, known as transcription, takes place in the cell nucleus.
But proteins are manufactured outside the cell nucleus, in microscopic structures called ribosomes. The mRNA is the vital link: the “courier” that picks up the protein-making instructions from DNA and delivers them to the protein-producing ribosomes.
RNA interference involves introducing miRNAs or siRNAs to a cell where they target, bind to and destroy specific mRNAs—effectively shutting down production of particular proteins and whatever gene expression they trigger or suppress.
Glossary
RNA—the acronym for ribonucleic acid, one of two nucleic acids found in the cells of every organism (the other is deoxyribonucleic acid, or DNA). RNA is composed of tiny molecules called nucleotides that are arranged in a chain. RNA transports the information needed to make proteins.
non-coding RNA (ncRNA)—a functional RNA molecule that is not translated into a protein. Examples of ncRNAs are short interfering RNAs and microRNAs.
short interfering RNAs (siRNAs)—small, double-stranded molecules that are used in RNA interference to prevent the translation of genes into proteins. siRNAs also are called small interfering RNAs or silencing RNAs.
microRNAs (miRNA)—small, hairpin-like structures of single-stranded RNA molecules that can regulate the expression of protein-coding genes.
RNA interference (RNAi)—a revolutionary technique for gene silencing. In this technique, RNA is introduced into a cell to disrupt messenger RNA and prevent it from being translated into a protein.
Services
The Center for RNA Interference and Non-Coding RNAs (RNA Center) is now providing high-throughput in situ hybridization (ISH) technology for microRNA and non-coding RNA analysis using Ventana Discovery Ultra. The Core provides equipment and services to determine gene expression patterns in mouse and human tissues.
Equipment:
- VENTANA DISCOVERY ULTRA
Flexibility:
- Unmatched protocol flexibility for rapid assay development &
- Run 30 completely independent protocols for improved workflow
Reproducibility:
- Fully automated slide processing, baking through staining
- Ready-to-use optimized reagents
- Apply precise heating to each slide
Materials and Methods:
- Frozen and FFPE tissue samples
- microRNA probe or custom LNA probe from Exiqon
- other ncRNA specific probes with LNA technology
- Detection with anti-digoxigenin or anti-FITC antibody /AP conjugated 2nd antibody/ NBT-BCIP substrate
- Imaging in a bright-field microscope
- Data interpretations are at the discretion of the investigator (we will provide a quantification of the intensity of the signal by using Leica Application Suite)
Processing Time:
- Up to 7 business days for miRNA hybridization
- Up to 10 business days for other ncRNA specific hybridization
At this stage, the RNA Center can provide chromogenic ISH service. In the near future, we will also provide fluorescence based-ISH and multiple ISH/IHC.
For additional information, including pricing and submitting a request, please contact Xinna Zhang, Ph.D., Assistant Professor, RNA Center at xzhang12@mdanderson.org.