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CRISPR Cas13-based tools to track and manipulate endogenous telomeric repeat-containing RNAs in living cells

Meng Xu, Tafadzwa Chigumira, Ziheng Chen, Jason Tones, Rongwei Zhao, Kris Noel Dahl, David M. Chenoweth, Huaiying Zhang
doi: https://doi.org/10.1101/2021.12.03.471109
Meng Xu
1Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University, Pittsburgh, PA, United States
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Tafadzwa Chigumira
2Department of Chemical Engineering, College of Engineering, Carnegie Mellon University, Pittsburgh, PA, United States
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Ziheng Chen
1Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University, Pittsburgh, PA, United States
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Jason Tones
1Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University, Pittsburgh, PA, United States
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Rongwei Zhao
1Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University, Pittsburgh, PA, United States
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Kris Noel Dahl
2Department of Chemical Engineering, College of Engineering, Carnegie Mellon University, Pittsburgh, PA, United States
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David M. Chenoweth
3Department of Chemistry, University of Pennsylvania, Philadelphia, PA, United States
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Huaiying Zhang
1Department of Biological Sciences, Mellon College of Science, Carnegie Mellon University, Pittsburgh, PA, United States
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  • For correspondence: huaiyinz@andrew.cmu.edu
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Abstract

TERRA, TElomeric Repeat-containing RNA, is a long non-coding RNA transcribed from telomeres. Emerging evidence indicates that TERRA regulates telomere maintenance and chromosome end protection in normal and cancerous cells. However, the mechanism of how TERRA contributes to telomere functions is still unclear, partially owing to the shortage of approaches to track and manipulate endogenous TERRA molecules in live cells. Here, we developed a method to visualize TERRA in live cells via a combination of CRISPR Cas13 RNA labeling and Suntag technology. Single-particle tracking reveals that TERRA foci undergo anomalous diffusion in a manner that depends on the timescale and telomeric localization. Furthermore, we used a chemically-induced protein dimerization system to manipulate TERRA subcellular localization in live cells. Overall, our approaches to monitor and control TERRA locations in live cells provide powerful tools to better understand its roles in telomere maintenance and genomic integrity.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 04, 2021.
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CRISPR Cas13-based tools to track and manipulate endogenous telomeric repeat-containing RNAs in living cells
Meng Xu, Tafadzwa Chigumira, Ziheng Chen, Jason Tones, Rongwei Zhao, Kris Noel Dahl, David M. Chenoweth, Huaiying Zhang
bioRxiv 2021.12.03.471109; doi: https://doi.org/10.1101/2021.12.03.471109
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CRISPR Cas13-based tools to track and manipulate endogenous telomeric repeat-containing RNAs in living cells
Meng Xu, Tafadzwa Chigumira, Ziheng Chen, Jason Tones, Rongwei Zhao, Kris Noel Dahl, David M. Chenoweth, Huaiying Zhang
bioRxiv 2021.12.03.471109; doi: https://doi.org/10.1101/2021.12.03.471109

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