Transient and Heritable Zebrafish Genetic Manipulations
Morpholino oligonucleotides (MOs) used for microinjection were ordered from Gene Tools. The MOs were microinjected into one-cell-stage embryos as previously described (Nasevicius and Ekker, 2000 (link)). Transient tprb transgene construct within Tol2 vectors (40 pg) was microinjected into one-cell-stage embryos with Tol2 transposase mRNA (50 pg). CRISPR-Cas9–mediated generation of zebrafish mutants (tprb–3+1 and vhl) were performed as previously described (Xiao et al., 2013 (link)). The gRNAs (50 pg) and Cas9 protein (500 pg) were co-microinjected into one-cell-stage embryos.
Wu S., Chen K., Xu T., Ma K., Gao L., Fu C., Zhang W., Jing C., Ren C., Deng M., Chen Y., Zhou Y., Pan W, & Jia X. (2021). Tpr Deficiency Disrupts Erythroid Maturation With Impaired Chromatin Condensation in Zebrafish Embryogenesis. Frontiers in Cell and Developmental Biology, 9, 709923.
Other organizations :
Chinese Academy of Sciences, Shanghai Institute of Nutrition and Health, Qingdao Agricultural University, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai Institute of Hematology, Boston Children's Hospital, Harvard University, Baotou Medical College
Transient tprb transgene construct within Tol2 vectors (40 pg)
CRISPR-Cas9 gRNAs (50 pg) and Cas9 protein (500 pg)
dependent variables
Phenotype/effects of MO knockdown
Phenotype/effects of tprb transgene expression
Phenotype/effects of tprb and vhl gene mutations
control variables
Microinjection of one-cell-stage embryos
Tol2 transposase mRNA (50 pg) co-injected with tprb transgene construct
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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