The Fusion of CLEC12A and MIR223HG Arises from a trans-Splicing Event in Normal and Transformed Human Cells.

Bijay P Dhungel,Mehdi S Tabar,Ulf Schmitz,Angela R M Fontaine,John E J Rasko,Geoffray Monteuuis,Divya Gokal,Jaynish S Shah,Caroline Giardina,Yue Feng,Cynthia Metierre,Sarah Ho,Rajini Nagarajah,Charles G Bailey

doi:10.3390/ijms222212178

Bijay P Dhungel, Mehdi S Tabar + Show 12 more

Open Access

PDF Available

https://doi.org/10.3390/ijms222212178

Copy DOI

Export

Save

Cite

Abstract
Highlights/Summary
Full-Text PDF
Similar Papers

Abstract

Listen

Chimeric RNAs are often associated with chromosomal rearrangements in cancer. In addition, they are also widely detected in normal tissues, contributing to transcriptomic complexity. Despite their prevalence, little is known about the characteristics and functions of chimeric RNAs. Here, we examine the genetic structure and biological roles of CLEC12A-MIR223HG, a novel chimeric transcript produced by the fusion of the cell surface receptor CLEC12A and the miRNA-223 host gene (MIR223HG), first identified in chronic myeloid leukemia (CML) patients. Surprisingly, we observed that CLEC12A-MIR223HG is not just expressed in CML, but also in a variety of normal tissues and cell lines. CLEC12A-MIR223HG expression is elevated in pro-monocytic cells resistant to chemotherapy and during monocyte-to-macrophage differentiation. We observed that CLEC12A-MIR223HG is a product of trans-splicing rather than a chromosomal rearrangement and that transcriptional activation of CLEC12A with the CRISPR/Cas9 Synergistic Activation Mediator (SAM) system increases CLEC12A-MIR223HG expression. CLEC12A-MIR223HG translates into a chimeric protein, which largely resembles CLEC12A but harbours an altered C-type lectin domain altering key disulphide bonds. These alterations result in differences in post-translational modifications, cellular localization, and protein–protein interactions. Taken together, our observations support a possible involvement of CLEC12A-MIR223HG in the regulation of CLEC12A function. Our workflow also serves as a template to study other uncharacterized chimeric RNAs.

Highlights

IntroductionChimeric RNAs are transcripts that consist of exons from different parental genes
Chimeric RNAs are transcripts that consist of exons from different parental genes.They can be produced by several mechanisms which may or may not involve chromosomal translocations at the genomic level
We examined the genetic architecture and the potential fusion protein encoded by the CLEC12A-MIR223HG chimeric transcript

Summary

Introduction

Chimeric RNAs are transcripts that consist of exons from different parental genes They can be produced by several mechanisms which may or may not involve chromosomal translocations at the genomic level. Transcription of fusion genes resulting from chromosomal deletion, inversion, or translocation are considered to be a hallmark of cancer [1,2] These fusion genes can give rise to proteins with important roles in cancer development and progression and may serve as therapeutic targets. Apart from chimeric RNAs produced by DNA-level gene fusions, studies have uncovered additional mechanisms that are known to produce chimeric transcripts at the RNA level [1,6,7,8] These include intergenic splicing by either cis-splicing, which involves same-strand neighbouring genes or by trans-splicing, whereby exons from two separate RNA transcripts are spliced together [1,9,10]. Cis- and trans- splicing events were originally thought to be rare in mammals, high-throughput analyses of transcriptomes have revealed that significant portions of chimeric RNAs are derived from intergenic splicing [11,12,13]

Methods

Results

Conclusion