Ichino, Noriko ORCID: 0000-0002-7009-8299, Serres, MaKayla R., Urban, Rhianna M., Urban, Mark D., Treichel, Anthony J., Schaefbauer, Kyle J., Greif, Lauren E., Varshney, Gaurav K. ORCID: 0000-0002-0429-1904, Skuster, Kimberly J., McNulty, Melissa S., Daby, Camden L., Wang, Ying, Liao, Hsin-kai, El-Rass, Suzan, Ding, Yonghe, Liu, Weibin, Anderson, Jennifer L., Wishman, Mark D., Sabharwal, Ankit, Schimmenti, Lisa A., Sivasubbu, Sridhar, Balciunas, Darius, Hammerschmidt, Matthias, Farber, Steven Arthur, Wen, Xiao-Yan, Xu, Xiaolei, McGrail, Maura ORCID: 0000-0001-9308-6189, Essner, Jeffrey J., Burgess, Shawn M., Clark, Karl J. and Ekker, Stephen C. (2020). Building the vertebrate codex using the gene breaking protein trap library. eLife, 9. CAMBRIDGE: ELIFE SCIENCES PUBLICATIONS LTD. ISSN 2050-084X

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Abstract

One key bottleneck in understanding the human genome is the relative under-characterization of 90% of protein coding regions. We report a collection of 1200 transgenic zebrafish strains made with the gene-break transposon (GBT) protein trap to simultaneously report and reversibly knockdown the tagged genes. Protein trap-associated mRFP expression shows previously undocumented expression of 35% and 90% of cloned genes at 2 and 4 days post-fertilization, respectively. Further, investigated alleles regularly show 99% gene-specific mRNA knockdown. Homozygous GBT animals in ryr1b, fras1, tnnt2a, edar and hmcn1 phenocopied established mutants. 204 cloned lines trapped diverse proteins, including 64 orthologs of human disease-associated genes with 40 as potential new disease models. Severely reduced skeletal muscle Ca2+ transients in GBT ryr1b homozygous animals validated the ability to explore molecular mechanisms of genetic diseases. This GBT system facilitates novel functional genome annotation towards understanding cellular and molecular underpinnings of vertebrate biology and human disease.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Ichino, NorikoUNSPECIFIEDorcid.org/0000-0002-7009-8299UNSPECIFIED
Serres, MaKayla R.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Urban, Rhianna M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Urban, Mark D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Treichel, Anthony J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schaefbauer, Kyle J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Greif, Lauren E.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Varshney, Gaurav K.UNSPECIFIEDorcid.org/0000-0002-0429-1904UNSPECIFIED
Skuster, Kimberly J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
McNulty, Melissa S.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Daby, Camden L.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wang, YingUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Liao, Hsin-kaiUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
El-Rass, SuzanUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ding, YongheUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Liu, WeibinUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Anderson, Jennifer L.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wishman, Mark D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Sabharwal, AnkitUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schimmenti, Lisa A.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Sivasubbu, SridharUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Balciunas, DariusUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hammerschmidt, MatthiasUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Farber, Steven ArthurUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wen, Xiao-YanUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Xu, XiaoleiUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
McGrail, MauraUNSPECIFIEDorcid.org/0000-0001-9308-6189UNSPECIFIED
Essner, Jeffrey J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Burgess, Shawn M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Clark, Karl J.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ekker, Stephen C.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-323458
DOI: 10.7554/eLife.54572
Journal or Publication Title: eLife
Volume: 9
Date: 2020
Publisher: ELIFE SCIENCES PUBLICATIONS LTD
Place of Publication: CAMBRIDGE
ISSN: 2050-084X
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
INSERTIONAL MUTAGENESIS; GENOME-WIDE; ZEBRAFISH; EXPRESSION; MODEL; IDENTIFICATION; VISUALIZATION; ACCUMULATION; MUTATIONS; SEQUENCEMultiple languages
BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/32345

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