Abstract
The most direct approach to study the physiological role of a protein of unknown function (Functional Genomics) is to change its expression pattern in an intact organism and analyze the phenotypic consequences of this manipulation. The introduction of a method to generate stably transgenic Xenopus laevis has paved the way to the use of tissue / cell- and developmental stage-specific promoters allowing to study the physiological function of proteins in a defined set of fully differentiated cells. Whereas stable (over)expression of proteins in Xenopus is now within reach, stable inhibition of protein expression can only be accomplished randomly, by gene trap approaches. We here report our efforts to induce stable RNA interference (RNAi) in X. laevis via transgene-driven expression of inverted repeats. Stable, and muscle- and neuron-specific knock-down of expression of exogenous green fluorescent protein (GFP) reporter was achieved via RNA polymerase II promoter-driven expression of long GFP RNA duplexes. Unfortunately, our attempts to induce RNAi directed against various endogenous targets, based on the use of RNA polymerase II and III promoters, and long and short inverted repeats have not resulted in a reliable protocol for stable, transgene-driven RNAi in Xenopus. In the second part, we present an example of the use of a cell-specific promoter for functional studies. Cell-specific transgene overexpression of a GFP-tagged member of the p24 family thought to be involved in intracellular protein transport was achieved and this manipulation of the intermediate pituitary melanotrope cell had a phenotypic consequence at its physiological target, the skin melanophore. Thus, the traditional experimental advantages of X. laevis combined with the recently developed technique of stable, non-mosaic Xenopus transgenesis make this lower vertebrate an attractive model organism for Functional Genomics.
Keywords: dsrna, intermediate pituitary cells, p24
Current Genomics
Title: Functional Genomics in Xenopus laevis: Towards Transgene-Driven RNA Interference and Cell-Specific Transgene Expression
Volume: 4 Issue: 8
Author(s): Ron P.H. Dirks, Gerrit Bouw, Rick Van Huizen, Eric J.R. Jansen and Gerard J.M. Martens
Affiliation:
Keywords: dsrna, intermediate pituitary cells, p24
Abstract: The most direct approach to study the physiological role of a protein of unknown function (Functional Genomics) is to change its expression pattern in an intact organism and analyze the phenotypic consequences of this manipulation. The introduction of a method to generate stably transgenic Xenopus laevis has paved the way to the use of tissue / cell- and developmental stage-specific promoters allowing to study the physiological function of proteins in a defined set of fully differentiated cells. Whereas stable (over)expression of proteins in Xenopus is now within reach, stable inhibition of protein expression can only be accomplished randomly, by gene trap approaches. We here report our efforts to induce stable RNA interference (RNAi) in X. laevis via transgene-driven expression of inverted repeats. Stable, and muscle- and neuron-specific knock-down of expression of exogenous green fluorescent protein (GFP) reporter was achieved via RNA polymerase II promoter-driven expression of long GFP RNA duplexes. Unfortunately, our attempts to induce RNAi directed against various endogenous targets, based on the use of RNA polymerase II and III promoters, and long and short inverted repeats have not resulted in a reliable protocol for stable, transgene-driven RNAi in Xenopus. In the second part, we present an example of the use of a cell-specific promoter for functional studies. Cell-specific transgene overexpression of a GFP-tagged member of the p24 family thought to be involved in intracellular protein transport was achieved and this manipulation of the intermediate pituitary melanotrope cell had a phenotypic consequence at its physiological target, the skin melanophore. Thus, the traditional experimental advantages of X. laevis combined with the recently developed technique of stable, non-mosaic Xenopus transgenesis make this lower vertebrate an attractive model organism for Functional Genomics.
Export Options
About this article
Cite this article as:
Dirks P.H. Ron, Bouw Gerrit, Huizen Van Rick, Jansen J.R. Eric and Martens J.M. Gerard, Functional Genomics in Xenopus laevis: Towards Transgene-Driven RNA Interference and Cell-Specific Transgene Expression, Current Genomics 2003; 4 (8) . https://dx.doi.org/10.2174/1389202033490132
DOI https://dx.doi.org/10.2174/1389202033490132 |
Print ISSN 1389-2029 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5488 |
Call for Papers in Thematic Issues
Advanced AI Techniques in Big Genomic Data Analysis
The thematic issue on "Advanced AI Techniques in Big Genomic Data Analysis" aims to explore the cutting-edge methodologies and applications of artificial intelligence (AI) in the realm of genomic research, where vast amounts of data pose both challenges and opportunities. This issue will cover a broad spectrum of AI-driven strategies, ...read more
Advanced Computational Algorithms and Artificial Intelligence in Clinical Pharmacogenomics
In the era of personalized medicine, understanding the relationship between genetics and drug response is crucial. This issue delves into innovative methodologies, leveraging deep computational analysis and artificial intelligence, to enhance the field of Clinical Pharmacogenomics. The interdisciplinary approach harnesses the power of advanced high-throughput genotyping technologies, sophisticated computational analysis, ...read more
Applications of Single-cell Sequencing Technology in Reproductive Medicine
Single cell sequencing (SCS) technology utilizes individual cells' genetic material to sequence their genome, transcriptome, and epigenetics at the molecular level. It offers insights into cell heterogeneity and enables the study of limited biological materials. Since its recognition as a valuable technique in 2011, single cell sequencing has yielded numerous ...read more
Big Data in Cancer Research
Cancer is a significant threat to human life and health, remaining a highly aggressive killer. It is a leading cause of death worldwide and represents a crucial medical issue for humanity. However, in the past decade, the effectiveness of new synthetic anticancer agents has not matched the current clinical speculation. ...read more
Related Journals
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Preface
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Diabetic Complications and Insight into Antidiabetic Potentialities of Ethno- Medicinal Plants: A Review
Recent Patents on Inflammation & Allergy Drug Discovery A Scientific Approach to Anti-Ageing Therapies: State of the Art
Current Pharmaceutical Design Epigenetic Modifications: New Therapeutic Targets in Primary Myelofibrosis
Current Stem Cell Research & Therapy Therapeutic Cancer Vaccines: At Midway Between Immunology and Pharmacology
Current Cancer Drug Targets Eotaxins and CCR3 Receptor in Inflammatory and Allergic Skin Diseases: Therapeutical Implications
Current Drug Targets - Inflammation & Allergy Tailoring Antiplatelet Therapy: A Step Toward Individualized Therapy to Improve Clinical Outcome?
Current Pharmaceutical Design Differential Regulation of Protein- and Polysaccharide-Specific Ig Isotype Production In Vivo in Response to Intact Streptococcus pneumoniae
Current Protein & Peptide Science New Drugs for Follicular Lymphoma in Older Adults
Anti-Cancer Agents in Medicinal Chemistry Prevalence and Predictors of Self-Medication Practices in India: A Systematic Literature Review and Meta-Analysis
Current Clinical Pharmacology Toxicogenomics of Cationic Lipid-based Vectors for Gene Therapy: Impact of Microarray Technology
Current Drug Delivery Use of Primary Macrophages for Searching Novel Immunocorrectors
Current Pharmaceutical Design Dietary Intake and Bone Status with Aging
Current Pharmaceutical Design Decoding the Mechanism of Drugs of Heterocyclic Nature against Hepatocellular Carcinoma
Anti-Cancer Agents in Medicinal Chemistry The Concept of Protein Mosaics: Physiological Role and Relevance for Prion Disease
Current Proteomics Patent Selections
Recent Patents on DNA & Gene Sequences Recent Patents on Rho Signaling Pathway as Therapeutic Target for Cardiovascular Diseases
Recent Patents on Cardiovascular Drug Discovery Antiangiogenic Therapy and Ovarian Cancer
Current Women`s Health Reviews Drug Tissue Distribution: Study Methods and Therapeutic Implications
Current Pharmaceutical Design Hibiscus Sabdariffa Linnaeus (Malvaceae), Curcumin and Resveratrol as Alternative Medicinal Agents Against Metabolic Syndrome
Cardiovascular & Hematological Agents in Medicinal Chemistry