Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

Conditional, tissue-specific CRISPR/Cas9 vector system in zebrafish reveal the role of neuropilin-1b in heart regeneration

Ramcharan Singh Angom, Ying Wang, Enfeng Wang, Shamit Dutta, Debabrata Mukhopadhyay
doi: https://doi.org/10.1101/2023.01.16.524222
Ramcharan Singh Angom
1Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Jacksonville, FL 32224
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Ying Wang
2Department of Cardiovascular Medicine, Mayo Clinic, Rochester, MN 55905
3Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Rochester, MN 55905
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Enfeng Wang
1Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Jacksonville, FL 32224
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Shamit Dutta
1Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Jacksonville, FL 32224
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Debabrata Mukhopadhyay
1Department of Biochemistry and Molecular Biology, College of Medicine and Science, Mayo Clinic, Jacksonville, FL 32224
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: Mukhopadhyay.debabrata@mayo.edu
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Supplementary material
  • Preview PDF
Loading

Abstract

CRISPR/Cas9 technology-mediated genome editing has significantly improved the targeted inactivation of genes in vitro and in vivo in many organisms. In this study, we have reported a novel CRISPR-based vector system for conditional tissue-specific gene ablation in zebrafish. Specifically, the cardiac-specific cardiac myosins light chain 2 (cmlc2) promoter drives Cas9 expression to silence the neuropilin-1(nrp1) gene in cardiomyocytes in a heat-shock inducible manner. This vector system establishes a unique tool to regulate the gene knockout in both the developmental and adult stages and hence, widens the possibility of loss-of-function studies in zebrafish at different stages of development and adulthood. Using this approach, we investigated the role of neuropilin isoforms nrp1a and nrp1b in response to cardiac injury and regeneration in adult zebrafish hearts. We observed that both the isoforms (nrp1a and nrp1b) are upregulated after the cryoinjury. Interestingly, the nrp1b-knockout significantly altered heart regeneration and impaired cardiac function in the adult zebrafish, demonstrated by reduced heart rate (ECG), ejection fractions, and fractional shortening. In addition, we show that the knockdown of nrp1b but not nrp1a induces activation of the cardiac remodeling genes in response to cryoinjury. To our knowledge, this is the first study where we have reported a heat shock-mediated conditional knockdown of nrp1a and nrp1b isoforms using CRISPR/Cas9 technology in the cardiomyocyte in zebrafish, and furthermore have identified a crucial role for nrp1b isoform in zebrafish cardiac remodeling and eventually heart function in response to injury.

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. All rights reserved. No reuse allowed without permission.
Back to top
PreviousNext
Posted January 17, 2023.
Download PDF

Supplementary Material

Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
Conditional, tissue-specific CRISPR/Cas9 vector system in zebrafish reveal the role of neuropilin-1b in heart regeneration
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Conditional, tissue-specific CRISPR/Cas9 vector system in zebrafish reveal the role of neuropilin-1b in heart regeneration
Ramcharan Singh Angom, Ying Wang, Enfeng Wang, Shamit Dutta, Debabrata Mukhopadhyay
bioRxiv 2023.01.16.524222; doi: https://doi.org/10.1101/2023.01.16.524222
Reddit logo Twitter logo Facebook logo LinkedIn logo Mendeley logo
Citation Tools
Conditional, tissue-specific CRISPR/Cas9 vector system in zebrafish reveal the role of neuropilin-1b in heart regeneration
Ramcharan Singh Angom, Ying Wang, Enfeng Wang, Shamit Dutta, Debabrata Mukhopadhyay
bioRxiv 2023.01.16.524222; doi: https://doi.org/10.1101/2023.01.16.524222

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Molecular Biology
Subject Areas
All Articles
  • Animal Behavior and Cognition (4237)
  • Biochemistry (9147)
  • Bioengineering (6786)
  • Bioinformatics (24025)
  • Biophysics (12137)
  • Cancer Biology (9545)
  • Cell Biology (13795)
  • Clinical Trials (138)
  • Developmental Biology (7642)
  • Ecology (11716)
  • Epidemiology (2066)
  • Evolutionary Biology (15518)
  • Genetics (10650)
  • Genomics (14332)
  • Immunology (9493)
  • Microbiology (22858)
  • Molecular Biology (9103)
  • Neuroscience (49032)
  • Paleontology (355)
  • Pathology (1484)
  • Pharmacology and Toxicology (2572)
  • Physiology (3849)
  • Plant Biology (8338)
  • Scientific Communication and Education (1472)
  • Synthetic Biology (2296)
  • Systems Biology (6196)
  • Zoology (1302)