1. Building the vertebrate codex using the gene breaking protein trap library.
- Author
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Ichino N, Serres MR, Urban RM, Urban MD, Treichel AJ, Schaefbauer KJ, Tallant LE, Varshney GK, Skuster KJ, McNulty MS, Daby CL, Wang Y, Liao HK, El-Rass S, Ding Y, Liu W, Anderson JL, Wishman MD, Sabharwal A, Schimmenti LA, Sivasubbu S, Balciunas D, Hammerschmidt M, Farber SA, Wen XY, Xu X, McGrail M, Essner JJ, Burgess SM, Clark KJ, and Ekker SC
- Subjects
- Animals, Animals, Genetically Modified genetics, Animals, Genetically Modified metabolism, RNA, Messenger genetics, Zebrafish metabolism, Zebrafish Proteins metabolism, Gene Knockdown Techniques, Gene Library, Genes, Reporter, Zebrafish genetics, Zebrafish Proteins genetics
- 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 Ca
2+ 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., Competing Interests: NI, MS, RU, MU, AT, KS, LG, GV, KS, MM, CD, YW, HL, SE, YD, WL, JA, MW, AS, LS, SS, DB, MH, SF, XW, XX, MM, JE, SB, KC, SE No competing interests declared- Published
- 2020
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