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Template plasmid integration in germline genome-edited cattle

View ORCID ProfileAlexis L. Norris, View ORCID ProfileStella S. Lee, Kevin J. Greenlees, Daniel A. Tadesse, Mayumi F. Miller, Heather Lombardi
doi: https://doi.org/10.1101/715482
Alexis L. Norris
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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Stella S. Lee
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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Kevin J. Greenlees
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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Daniel A. Tadesse
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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Mayumi F. Miller
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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Heather Lombardi
Center for Veterinary Medicine, Food and Drug Administration, Rockville, MD 20855
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  • For correspondence: Heather.Lombardi@fda.hhs.gov
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Abstract

We analyzed publicly available whole genome sequencing data from cattle which were germline genome-edited to introduce polledness. Our analysis discovered the unintended heterozygous integration of the plasmid and a second copy of the repair template sequence, at the target site. Our finding underscores the importance of employing screening methods suited to reliably detect the unintended integration of plasmids and multiple template copies.

Footnotes

  • Competing Financial Interests: The authors declare no competing financial interests.

  • https://trace.ncbi.nlm.nih.gov/Traces/sra/?study=SRP072240

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. This article is a US Government work. It is not subject to copyright under 17 USC 105 and is also made available for use under a CC0 license.
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Posted July 28, 2019.
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Template plasmid integration in germline genome-edited cattle
Alexis L. Norris, Stella S. Lee, Kevin J. Greenlees, Daniel A. Tadesse, Mayumi F. Miller, Heather Lombardi
bioRxiv 715482; doi: https://doi.org/10.1101/715482
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Template plasmid integration in germline genome-edited cattle
Alexis L. Norris, Stella S. Lee, Kevin J. Greenlees, Daniel A. Tadesse, Mayumi F. Miller, Heather Lombardi
bioRxiv 715482; doi: https://doi.org/10.1101/715482

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