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In situ Nuclear Matrix preparation in Drosophila melanogaster and its use in studying the components of nuclear architecture

Rashmi U Pathak, Rahul Sureka, Ashish Bihani, Parul Varma, View ORCID ProfileRakesh K Mishra
doi: https://doi.org/10.1101/2021.09.30.462611
Rashmi U Pathak
1Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500 007, India
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Rahul Sureka
2EMBL, Rome, Italy
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Ashish Bihani
1Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500 007, India
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Parul Varma
3Department of Neuroscience, Development and Regenerative Biology, The University of Texas at San Antonio, Texas, USA
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Rakesh K Mishra
1Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad-500 007, India
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  • ORCID record for Rakesh K Mishra
  • For correspondence: mishra@ccmb.res.in
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Abstract

The study of Nuclear Matrix (NuMat) over the last 40 years has been limited to either isolated nuclei from tissues or cells grown in culture. Here, we provide a protocol for NuMat preparation in intact Drosophila melanogaster embryos and its use in dissecting the components of nuclear architecture. The protocol does not require isolation of nuclei and therefore maintains the three-dimensional milieu of an intact embryo, which is biologically more relevant compared to cells in culture. One of the advantages of this protocol is that only a small number of embryos are required. The protocol can be extended to larval tissues like salivary glands and imaginal discs with little modification. Taken together, it becomes possible to carry out such studies in parallel to genetic experiments using mutant and transgenic flies. This protocol, therefore, opens the powerful field of fly genetics to cell biology in the study of nuclear architecture.

Summary Nuclear Matrix is a biochemically defined entity and a basic component of the nuclear architecture. Here we present a protocol to isolate and visualize Nuclear Matrix in situ in the intact embryos and tissues of Drosophila melanogaster and its potential applications.

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 4.0 International license.
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Posted September 30, 2021.
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In situ Nuclear Matrix preparation in Drosophila melanogaster and its use in studying the components of nuclear architecture
Rashmi U Pathak, Rahul Sureka, Ashish Bihani, Parul Varma, Rakesh K Mishra
bioRxiv 2021.09.30.462611; doi: https://doi.org/10.1101/2021.09.30.462611
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In situ Nuclear Matrix preparation in Drosophila melanogaster and its use in studying the components of nuclear architecture
Rashmi U Pathak, Rahul Sureka, Ashish Bihani, Parul Varma, Rakesh K Mishra
bioRxiv 2021.09.30.462611; doi: https://doi.org/10.1101/2021.09.30.462611

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