12. Overexpression of Chimeric RNA by Retroviral Transduction
Hao Wu and Hui Li
13. Separation of Nuclear and Cytoplasmic Fractions for Chimeric RNA Characterization
Xinrui Shi, Fujun Qin, and Hui Li
14. Confirmation of Transcriptional Read-Through Events by RT-PCR
Xinrui Shi, Fujun Qin, and Hui Li
15. Validating Gene Fusion as the Source of Chimeric RNAs
Sachin Kumar Gupta, Jocelyn Duen-Ya Jea, and Laising Yen
Part IV: Translational Applications
16. Chimeric RNA and Exosomes-Based Liquid Biopsy
Xiurong Ke, Xiao Xiong, Yusheng Lin, and Hao Zhang
17. SMaRT for Therapeutic Purposes
Lisa M. Riedmayr
18. Case Study: Landscape of Chimeric RNAs in Bladder Cancer
Dingjun Zhu and Hui Li
19. Case Study: The Recurrent Fusion RNA DUS4L-BCAP29 in Non-Cancer Human Tissues and Cells
Yue Tang, Fangxia Guan, and Hui Li
This book provides numerous methods for identification, validation, and functional characterization of chimeric RNAs, herein described as any transcript which contains the nucleotide sequence of two distinct parental genes. Beginning with identification and characterization through bioinformatics tools, the book continues with experimental validation using RT-PCR, hybridization methods for RNA, and the use of mass spectrometry data for fusion protein validation, as well as functional and mechanistic studies of chimeric RNAs and translational applications. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Chimeric RNA: Methods and Protocols is an ideal guide to this expansive field and an inspiration to researchers seeking to further our knowledge of these remarkable features of the transcriptome.