mRNA PCR-Based Epitope Chase Method
The identification of specific viral and tumor antigen epitopes recognized by CD4+ or CD8+ T lymphocytes remains a challenge. Unfortunately, epitope mapping methods are generally costly and time-consuming. This chapter details a polymerase chain reaction (PCR)-based mRNA epitope identification method called mPEC, which is designed to rapidly and precisely identify relevant T cell epitopes recognized by previously isolated CD8+ or CD4+ T lymphocytes. This method is based on the use of mRNA fragments synthesized from PCR-amplified cDNA with a variety of 3′end iterative deletions. mRNA fragments are electroporated into autologous antigen-presenting cells to map the epitope in a given protein antigen. Considering mRNA’s sensitivity to degradation, we also insert a control define epitope at the mRNA’s 3′end to control for electroporated mRNA’s integrity and capacity to be translated.
- PRINS DNA Synthesis on Frozen Tissue Sections
- Methods for Selecting Effective siRNA Sequences by using Statistical and Clustering Techniques
- Profiling of Short RNAs Using Helicos Single-Molecule Sequencing
- Detection and Interpretation of Genomic Structural Variation in Mammals
- Quality Control in FISH as Part of a Laboratorys Quality Management System
- Airway Epithelia
- Nonspecific, Nested Suppression PCR Method for Isolation of Unknown Flanking DNA (Cold-Start Method)
- MicroRNA Biogenesis: Isolation and Characterization of the Microprocessor Complex
- Generation of Gene-Specific Mutated Rats Using Zinc-Finger Nucleases
- RNA Structure Prediction: An Overview of Methods