2.50
Hdl Handle:
http://hdl.handle.net/10755/157811
Type:
Presentation
Title:
RNA-Based Biological Methods: An Emerging Field
Abstract:
RNA-Based Biological Methods: An Emerging Field
Conference Sponsor:Western Institute of Nursing
Conference Year:2009
Author:Merkle, Carrie J., PhD, RN, FAAN
P.I. Institution Name:University of Arizona, College of Nursing
Title:Associate Professor
Contact Address:1305 N. Martin, PO Box 210203, Tucson, AZ, 85721, USA
Contact Telephone:520-626-6163
Co-Authors:David W. Montgomery, PhD, Research Associate Professor; Ida M. (Ki) Moore, DNSc, RN, FAAN, Professor; Sharon A. Lobert, PhD, RN, Professor
Purpose: The purpose of this paper is to review state-of-the-science and emerging biological methods that focus on mRNA measurement and manipulation. Examples of use and data that are related to cancer research from our laboratories will be presented and discussed. Introduction and Rationale: Although DNA is the central focus in the fields of genetics and genomics, recent advances have contributed to an explosion of investigative interest in RNA and how it regulates cell functions, can be used as a platform for assays, and can be used as a tool to "silence genes." These advances resulted from the advent of the complete sequencing of the genome coupled with advances in biotechnology including the polymerase chain reaction (PCR); microarray and "chip"-based technologies; fluorescent probes and other detection mechanisms; and automated, computerized equipment. It has become relatively easy to examine changes in the expression of multiple genes in cells and tissues by profiling mRNA expression. These assays permit one to identify specific cellular mechanisms that become activated or suppressed in response to various treatments or changes in physiological conditions. Additionally, RNA interference (RNAi) techniques, which are based on natural cellular defense mechanisms that protect cells from viral RNA harm, have become popular methods to examine the effects of silencing particular genes on cellular responses and behavior. Collectively, our laboratories have experience with these techniques in endeavors to assist in better understanding the consequences of cancer and its treatment. Methods: Originally we examined changes in gene expression using an array format with a chemiluminescence-based detection system. More recently our studies have employed the RT2 Profiler PCR arrays (SABioscience). These are supplied in 96- and 384- well formats (to measure expression of 84 or 370 genes, respectively) to examine specific pathway- and disease-related gene expression. After isolation of the RNA, the protocol involves a rapid and simple real-time PCR method. Data are analyzed using a downloadable template that is easy to use. These PCR arrays permit one to identify specific cellular mechanisms that become activated or suppressed in response to various treatments or changes in physiological conditions, such as changes in cells in response to chemotherapy. As for RNAi methodology, we are currently performing studies to examine changes in breast cancer metastasis and responses to chemotherapy when specific proteins in breast cancer cells are reduced or "knocked down" using short hairpin RNAs to silence genes. Issues encountered in these studies will be discussed. Conclusion: In addition to the DNA focus of genetic/genomic studies, RNA-based methods are emerging as invaluable techniques in scientific endeavors that pertain to the biology of disease, such as cancer, and disease treatment. Technological advances such as automated and easy to use equipment, as well as commercially-available kits and products, have enabled these RNA-based methods to be used in nursing investigations.
Repository Posting Date:
26-Oct-2011
Date of Publication:
17-Oct-2011
Sponsors:
Western Institute of Nursing

Full metadata record

DC FieldValue Language
dc.typePresentationen_GB
dc.titleRNA-Based Biological Methods: An Emerging Fielden_GB
dc.identifier.urihttp://hdl.handle.net/10755/157811-
dc.description.abstract<table><tr><td colspan="2" class="item-title">RNA-Based Biological Methods: An Emerging Field</td></tr><tr class="item-sponsor"><td class="label">Conference Sponsor:</td><td class="value">Western Institute of Nursing</td></tr><tr class="item-year"><td class="label">Conference Year:</td><td class="value">2009</td></tr><tr class="item-author"><td class="label">Author:</td><td class="value">Merkle, Carrie J., PhD, RN, FAAN</td></tr><tr class="item-institute"><td class="label">P.I. Institution Name:</td><td class="value">University of Arizona, College of Nursing</td></tr><tr class="item-author-title"><td class="label">Title:</td><td class="value">Associate Professor</td></tr><tr class="item-address"><td class="label">Contact Address:</td><td class="value">1305 N. Martin, PO Box 210203, Tucson, AZ, 85721, USA</td></tr><tr class="item-phone"><td class="label">Contact Telephone:</td><td class="value">520-626-6163</td></tr><tr class="item-email"><td class="label">Email:</td><td class="value">cmerkle@nursing.arizona.edu</td></tr><tr class="item-co-authors"><td class="label">Co-Authors:</td><td class="value">David W. Montgomery, PhD, Research Associate Professor; Ida M. (Ki) Moore, DNSc, RN, FAAN, Professor; Sharon A. Lobert, PhD, RN, Professor</td></tr><tr><td colspan="2" class="item-abstract">Purpose: The purpose of this paper is to review state-of-the-science and emerging biological methods that focus on mRNA measurement and manipulation. Examples of use and data that are related to cancer research from our laboratories will be presented and discussed. Introduction and Rationale: Although DNA is the central focus in the fields of genetics and genomics, recent advances have contributed to an explosion of investigative interest in RNA and how it regulates cell functions, can be used as a platform for assays, and can be used as a tool to &quot;silence genes.&quot; These advances resulted from the advent of the complete sequencing of the genome coupled with advances in biotechnology including the polymerase chain reaction (PCR); microarray and &quot;chip&quot;-based technologies; fluorescent probes and other detection mechanisms; and automated, computerized equipment. It has become relatively easy to examine changes in the expression of multiple genes in cells and tissues by profiling mRNA expression. These assays permit one to identify specific cellular mechanisms that become activated or suppressed in response to various treatments or changes in physiological conditions. Additionally, RNA interference (RNAi) techniques, which are based on natural cellular defense mechanisms that protect cells from viral RNA harm, have become popular methods to examine the effects of silencing particular genes on cellular responses and behavior. Collectively, our laboratories have experience with these techniques in endeavors to assist in better understanding the consequences of cancer and its treatment. Methods: Originally we examined changes in gene expression using an array format with a chemiluminescence-based detection system. More recently our studies have employed the RT2 Profiler PCR arrays (SABioscience). These are supplied in 96- and 384- well formats (to measure expression of 84 or 370 genes, respectively) to examine specific pathway- and disease-related gene expression. After isolation of the RNA, the protocol involves a rapid and simple real-time PCR method. Data are analyzed using a downloadable template that is easy to use. These PCR arrays permit one to identify specific cellular mechanisms that become activated or suppressed in response to various treatments or changes in physiological conditions, such as changes in cells in response to chemotherapy. As for RNAi methodology, we are currently performing studies to examine changes in breast cancer metastasis and responses to chemotherapy when specific proteins in breast cancer cells are reduced or &quot;knocked down&quot; using short hairpin RNAs to silence genes. Issues encountered in these studies will be discussed. Conclusion: In addition to the DNA focus of genetic/genomic studies, RNA-based methods are emerging as invaluable techniques in scientific endeavors that pertain to the biology of disease, such as cancer, and disease treatment. Technological advances such as automated and easy to use equipment, as well as commercially-available kits and products, have enabled these RNA-based methods to be used in nursing investigations.</td></tr></table>en_GB
dc.date.available2011-10-26T20:13:38Z-
dc.date.issued2011-10-17en_GB
dc.date.accessioned2011-10-26T20:13:38Z-
dc.description.sponsorshipWestern Institute of Nursingen_GB
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