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1000 Titel
  • Development and evaluation of a milk protein transcript depletion method for differential transcriptome analysis in mammary gland tissue
1000 Autor/in
  1. Brodhagen, Johanna |
  2. Weikard, Rosemarie |
  3. Thom, Ulrike |
  4. Heimes, Annika |
  5. Günther, Juliane |
  6. Hadlich, Frieder |
  7. Zerbe, Holm |
  8. Petzl, Wolfgang |
  9. Meyerholz, Marie M. |
  10. Hoedemaker, Martina |
  11. Schuberth, Hans-Joachim |
  12. Engelmann, Susanne |
  13. Kuehn, Christa |
1000 Erscheinungsjahr 2019
1000 LeibnizOpen
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2019-05-22
1000 Erschienen in
1000 Quellenangabe
  • 20:400
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2019
1000 Lizenz
1000 Verlagsversion
  • https://doi.org/10.1186/s12864-019-5781-3 |
  • https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6530097/ |
1000 Ergänzendes Material
  • https://bmcgenomics.biomedcentral.com/articles/10.1186/s12864-019-5781-3#Sec25 |
1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • BACKGROUND: In the mammary gland transcriptome of lactating dairy cows genes encoding milk proteins are highly abundant, which can impair the detection of lowly expressed transcripts and can bias the outcome in global transcriptome analyses. Therefore, the aim of this study was to develop and evaluate a method to deplete extremely highly expressed transcripts in mRNA from lactating mammary gland tissue. RESULTS: Selective RNA depletion was performed by hybridization of antisense oligonucleotides targeting genes encoding the caseins (CSN1S1, CSN1S2, CSN2 and CSN3) and whey proteins (LALBA and PAEP) within total RNA followed by RNase H-mediated elimination of the respective transcripts. The effect of the RNA depletion procedure was monitored by RNA sequencing analysis comparing depleted and non-depleted RNA samples from Escherichia coli (E. coli) challenged and non-challenged udder tissue of lactating cows in a proof of principle experiment. Using RNase H-mediated RNA depletion, the ratio of highly abundant milk protein gene transcripts was reduced in all depleted samples by an average of more than 50% compared to the non-depleted samples. Furthermore, the sensitivity for discovering transcripts with marginal expression levels and transcripts not yet annotated was improved. Finally, the sensitivity to detect significantly differentially expressed transcripts between non-challenged and challenged udder tissue was increased without leading to an inadvertent bias in the pathogen challenge-associated biological signaling pathway patterns. CONCLUSIONS: The implementation of selective RNase H-mediated RNA depletion of milk protein gene transcripts from the mammary gland transcriptome of lactating cows will be highly beneficial to establish comprehensive transcript catalogues of the tissue that better reflects its transcriptome complexity.
1000 Sacherschließung
lokal RNA depletion
lokal Lactation
lokal Mammary gland
lokal RNA-sequencing
lokal Mastitis
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
  1. https://frl.publisso.de/adhoc/uri/QnJvZGhhZ2VuLCBKb2hhbm5h|https://frl.publisso.de/adhoc/uri/V2Vpa2FyZCwgUm9zZW1hcmll|https://frl.publisso.de/adhoc/uri/VGhvbSwgVWxyaWtl|https://frl.publisso.de/adhoc/uri/SGVpbWVzLCBBbm5pa2E=|https://frl.publisso.de/adhoc/uri/R8O8bnRoZXIsIEp1bGlhbmU=|https://frl.publisso.de/adhoc/uri/SGFkbGljaCwgRnJpZWRlcg==|https://frl.publisso.de/adhoc/uri/WmVyYmUsIEhvbG0=|https://frl.publisso.de/adhoc/uri/UGV0emwsIFdvbGZnYW5n|https://frl.publisso.de/adhoc/uri/TWV5ZXJob2x6LCBNYXJpZSBNLg==|https://frl.publisso.de/adhoc/uri/SG9lZGVtYWtlciwgTWFydGluYQ==|https://frl.publisso.de/adhoc/uri/U2NodWJlcnRoLCBIYW5zLUpvYWNoaW0=|https://frl.publisso.de/adhoc/uri/RW5nZWxtYW5uLCBTdXNhbm5l|https://orcid.org/0000-0002-0216-424X
1000 Label
1000 Förderer
  1. Landwirtschaftliche Rentenbank (Deutschland) |
  2. Leibniz-Institut für Nutztierbiologie |
1000 Fördernummer
  1. 740718
  2. -
1000 Förderprogramm
  1. Spezialfond
  2. Open Access Fund
1000 Dateien
1000 Förderung
  1. 1000 joinedFunding-child
    1000 Förderer Landwirtschaftliche Rentenbank (Deutschland) |
    1000 Förderprogramm Spezialfond
    1000 Fördernummer 740718
  2. 1000 joinedFunding-child
    1000 Förderer Leibniz-Institut für Nutztierbiologie |
    1000 Förderprogramm Open Access Fund
    1000 Fördernummer -
1000 Objektart article
1000 Beschrieben durch
1000 @id frl:6416807.rdf
1000 Erstellt am 2019-10-15T15:43:07.869+0200
1000 Erstellt von 218
1000 beschreibt frl:6416807
1000 Bearbeitet von 25
1000 Zuletzt bearbeitet Wed Oct 30 14:43:50 CET 2019
1000 Objekt bearb. Wed Oct 30 14:43:26 CET 2019
1000 Vgl. frl:6416807
1000 Oai Id
  1. oai:frl.publisso.de:frl:6416807 |
1000 Sichtbarkeit Metadaten public
1000 Sichtbarkeit Daten public
1000 Gegenstand von

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