Abstract
Identifying gene function in specific cells is critical for understanding the processes that make cells unique. Several different methods are available to isolate actively transcribed RNA or actively translated RNA in specific cells at a chosen time point. Cell-specific mRNA isolation can be accomplished by the expression of transgenes in cells of interest, either directly from a specific promoter or using a modular system such as Gal4/UAS or Cre/lox. All of the methods described in this review, namely thiol-labeling of RNA (TU-tagging or RABT), TRAP (translating ribosome affinity purification) and INTACT (isolation of nuclei tagged in specific cell types), allow next generation sequencing, permitting the identification of enriched gene transcripts within the specific cell-type. We describe here the general concept of each method, include examples, evaluate possible problems related to each technique, and suggest the types of questions for which each method is best suited.
Keywords: 4tU-tagging, Gene expression profiling, INTACT, RNA-seq, Transcriptome, Translatome, TRAP.
Current Genomics
Title:Temporally and Spatially Restricted Gene Expression Profiling
Volume: 15 Issue: 4
Author(s): Alexandra Tallafuss, Philip Washbourne and John Postlethwait
Affiliation:
Keywords: 4tU-tagging, Gene expression profiling, INTACT, RNA-seq, Transcriptome, Translatome, TRAP.
Abstract: Identifying gene function in specific cells is critical for understanding the processes that make cells unique. Several different methods are available to isolate actively transcribed RNA or actively translated RNA in specific cells at a chosen time point. Cell-specific mRNA isolation can be accomplished by the expression of transgenes in cells of interest, either directly from a specific promoter or using a modular system such as Gal4/UAS or Cre/lox. All of the methods described in this review, namely thiol-labeling of RNA (TU-tagging or RABT), TRAP (translating ribosome affinity purification) and INTACT (isolation of nuclei tagged in specific cell types), allow next generation sequencing, permitting the identification of enriched gene transcripts within the specific cell-type. We describe here the general concept of each method, include examples, evaluate possible problems related to each technique, and suggest the types of questions for which each method is best suited.
Export Options
About this article
Cite this article as:
Tallafuss Alexandra, Washbourne Philip and Postlethwait John, Temporally and Spatially Restricted Gene Expression Profiling, Current Genomics 2014; 15 (4) . https://dx.doi.org/10.2174/1389202915666140602230106
DOI https://dx.doi.org/10.2174/1389202915666140602230106 |
Print ISSN 1389-2029 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5488 |
Call for Papers in Thematic Issues
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
Current Genomics in Cardiovascular Research
Cardiovascular diseases are the main cause of death in the world, in recent years we have had important advances in the interaction between cardiovascular disease and genomics. In this Research Topic, we intend for researchers to present their results with a focus on basic, translational and clinical investigations associated with ...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
-
Platelets in Rheumatic Diseases: Friend or Foe?
Current Pharmaceutical Design Chemotherapeutic Options for Colorectal Cancer Patients with Cardiovascular Diseases.
Reviews on Recent Clinical Trials Recent Patents in Diagnosis and Treatment for Inborn Errors of Metabolism
Recent Patents on Endocrine, Metabolic & Immune Drug Discovery Predicting Drug Metabolism Induction In Silico
Current Topics in Medicinal Chemistry Vitamin D Deficiency: Universal Risk Factor for Multifactorial Diseases?
Current Drug Targets Wide Pulse Pressure in the Elderly
Current Cardiology Reviews Analysis of Current Antifungal Agents and Their Targets within the Pneumocystis carinii Genome
Current Drug Targets Severe Neutropenia in a Renal Transplant Patient Suggesting an Interaction Between Mycophenolate and Fenofibrate
Current Drug Safety Old and New Drugs for Treatment of Stable Angina: New Anti-Anginal Drugs and Coronary Revascularization
Cardiovascular & Hematological Agents in Medicinal Chemistry Reactive Oxygen Species, Vascular Disease and Cardiovascular Surgery
Current Vascular Pharmacology Oral Antiplatelet Therapy for Acute Coronary Syndromes: Aspirin, P2Y12 Inhibition and Thrombin Receptor Antagonists
Current Drug Targets Protein Kinases and Associated Pathways in Pluripotent State and Lineage Differentiation
Current Stem Cell Research & Therapy HIV Infection and Myocardial Infarction
Current HIV Research Disturbed Tryptophan Metabolism in Cardiovascular Disease
Current Medicinal Chemistry Evaluation and Comparative Correlation of Abdominal Fat Related Parameters in Obese and Non-obese Groups Using Computed Tomography
Current Medical Imaging The J-shaped Association Between Alcohol Consumption and Breast Cancer: A Case-Control Study
Current Nutrition & Food Science Nutrition Communication in General Practice
Current Nutrition & Food Science Persistent Pulmonary Hypertension of the Newborn: Physiology, Hemodynamic Assessment and Novel Therapies
Current Pediatric Reviews Efficacy and Cardiovascular Safety of Antidiabetic Medications
Current Drug Safety Metabolism of HDL and its Regulation
Current Medicinal Chemistry