CRISPR/Cas9-Mediated Knockout of tnfaip1 in Zebrafish Plays a Role in Early Development

TNF α-induced protein 1 (TNFAIP1) was first identified in human umbilical vein endothelial cells and can be induced by tumor necrosis factor α (TNFα). Early studies have found that TNFAIP1 is involved in the development of many tumors and is closely associated with the neurological disorder Alzheimer’s disease. However, little is known about the expression pattern of TNFAIP1 under physiological conditions and its function during embryonic development. In this study, we used zebrafish as a model to illustrate the early developmental expression pattern of tnfaip1 and its role in early development. First, we examined the expression pattern of tnfaip1 during early zebrafish development using quantitative real-time PCR and whole mount in situ hybridization and found that tnfaip1 was highly expressed in early embryonic development and, subsequently, expression became localized to anterior embryonic structures. To investigate the function of tnfaip1 during early development, we constructed a model of a stably inherited tnfaip1 mutant using the CRISPR/Cas9 system. Tnfaip1 mutant embryos showed significant developmental delays as well as microcephaly and microphthalmia. At the same time, we found decreased expression of the neuronal marker genes tuba1b, neurod1, and ccnd1 in tnfaip1 mutants. Analysis of transcriptome sequencing data revealed altered expression of the embryonic development related genes dhx40, hspa13, tnfrsf19, nppa, lrp2b, hspb9, clul1, zbtb47a, cryba1a, and adgrg4a in the tnfaip1 mutants. These findings suggest an important role for tnfaip1 in the early development of zebrafish.

[1]  Brian S Muntean,et al.  Multiple potassium channel tetramerization domain (KCTD) family members interact with Gβγ, with effects on cAMP signaling , 2023, The Journal of biological chemistry.

[2]  Jingruo Zhang,et al.  Downregulation of TNFAIP1 alleviates OGD/R‑induced neuronal damage by suppressing Nrf2/GPX4‑mediated ferroptosis , 2022, Experimental and therapeutic medicine.

[3]  Li Wang,et al.  Epigenetic axis of SNHG19/miR-137/TNFAIP1 modulates amyloid beta peptide 25-35-induced SH-SY5Y cytotoxicity. , 2022, Epigenomics.

[4]  S. Oh,et al.  Human gut-microbiome-derived propionate coordinates proteasomal degradation via HECTD2 upregulation to target EHMT2 in colorectal cancer , 2022, The ISME Journal.

[5]  J. Wohlschlegel,et al.  Members of the KCTD family are major regulators of cAMP signaling , 2021, Proceedings of the National Academy of Sciences.

[6]  L. Jia,et al.  Cullin3-TNFAIP1 E3 Ligase Controls Inflammatory Response in Hepatocellular Carcinoma Cells via Ubiquitination of RhoB , 2021, Frontiers in Cell and Developmental Biology.

[7]  G. Aguirre,et al.  Candidate Genetic Modifiers for RPGR Retinal Degeneration , 2020, Investigative ophthalmology & visual science.

[8]  Shuanglin Xiang,et al.  TNFAIP1 Is Upregulated in APP/PS1 Mice and Promotes Apoptosis in SH-SY5Y Cells by Binding to RhoB , 2020, Journal of Molecular Neuroscience.

[9]  Xiang Hu,et al.  Tumor necrosis factor α-induced protein 1 as a novel tumor suppressor through selective downregulation of CSNK2B blocks nuclear factor-κB activation in hepatocellular carcinoma , 2020, EBioMedicine.

[10]  L. Maquat,et al.  Quality and quantity control of gene expression by nonsense-mediated mRNA decay , 2019, Nature Reviews Molecular Cell Biology.

[11]  Yan Shi,et al.  Effects of XIST/miR‐137 axis on neuropathic pain by targeting TNFAIP1 in a rat model , 2018, Journal of cellular physiology.

[12]  O. Mühlemann,et al.  Beyond quality control: The role of nonsense-mediated mRNA decay (NMD) in regulating gene expression. , 2017, Seminars in cell & developmental biology.

[13]  Jiewen Zhang,et al.  miR-137 attenuates Aβ-induced neurotoxicity through inactivation of NF-κB pathway by targeting TNFAIP1 in Neuro2a cells. , 2017, Biochemical and biophysical research communications.

[14]  Jian-feng Dong,et al.  MiR-424 Promotes Non-Small Cell Lung Cancer Progression and Metastasis through Regulating the Tumor Suppressor Gene TNFAIP1 , 2017, Cellular Physiology and Biochemistry.

[15]  S. Burgess,et al.  A high-throughput functional genomics workflow based on CRISPR/Cas9-mediated targeted mutagenesis in zebrafish , 2016, Nature Protocols.

[16]  Siwen Hu,et al.  Caudatin targets TNFAIP1/NF-κB and cytochrome c/caspase signaling to suppress tumor progression in human uterine cancer. , 2016, International journal of oncology.

[17]  Xiang Hu,et al.  TNFAIP1 contributes to the neurotoxicity induced by Aβ25–35 in Neuro2a cells , 2016, BMC Neuroscience.

[18]  Shan Shan Li,et al.  Bacurd1/Kctd13 and Bacurd2/Tnfaip1 are interacting partners to Rnd proteins which influence the long-term positioning and dendritic maturation of cerebral cortical neurons , 2016, Neural Development.

[19]  C. Croce,et al.  MicroRNA-224 promotes tumor progression in nonsmall cell lung cancer , 2015, Proceedings of the National Academy of Sciences.

[20]  Shan Shan Li,et al.  Bacurd2 is a novel interacting partner to Rnd2 which controls radial migration within the developing mammalian cerebral cortex , 2015, Neural Development.

[21]  J. Sebat,et al.  Spatiotemporal 16p11.2 Protein Network Implicates Cortical Late Mid-Fetal Brain Development and KCTD13-Cul3-RhoA Pathway in Psychiatric Diseases , 2015, Neuron.

[22]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[23]  Jianlin Zhou,et al.  KCTD10 Is Involved in the Cardiovascular System and Notch Signaling during Early Embryonic Development , 2014, PloS one.

[24]  Tessa G. Montague,et al.  Efficient Mutagenesis by Cas9 Protein-Mediated Oligonucleotide Insertion and Large-Scale Assessment of Single-Guide RNAs , 2014, PloS one.

[25]  Xiang Hu,et al.  KCTD10 is critical for heart and blood vessel development of zebrafish. , 2014, Acta biochimica et biophysica Sinica.

[26]  Shuo Lin,et al.  Kctd10 regulates heart morphogenesis by repressing the transcriptional activity of Tbx5a in zebrafish , 2014, Nature Communications.

[27]  Yaqian Xiang,et al.  The KCTD family of proteins: structure, function, disease relevance , 2013, Cell & Bioscience.

[28]  Yaqian Xiang,et al.  The KCTD family of proteins: structure, function, disease relevance , 2013, Cell & Bioscience.

[29]  Xiang Hu,et al.  MicroRNA-373 is upregulated and targets TNFAIP1 in human gastric cancer, contributing to tumorigenesis , 2013, Oncology letters.

[30]  Xiaofeng Li,et al.  microRNA-372 maintains oncogene characteristics by targeting TNFAIP1 and affects NFκB signaling in human gastric carcinoma cells. , 2013, International journal of oncology.

[31]  A. Reymond,et al.  KCTD13 is a major driver of mirrored neuroanatomical phenotypes of the 16p11.2 copy number variant , 2012, Nature.

[32]  Colin N. Dewey,et al.  RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome , 2011, BMC Bioinformatics.

[33]  Yeon-Soo Seo,et al.  RhoB induces apoptosis via direct interaction with TNFAIP1 in HeLa cells , 2009, International journal of cancer.

[34]  J. Luk,et al.  Tumor necrosis factor-α-induced protein 1 and immunity to hepatitis B virus , 2005 .

[35]  Xiaoxiao Hu,et al.  Cloning of two rat PDIP1 related genes and their interactions with proliferating cell nuclear antigen. , 2005, Journal of experimental zoology. Part A, Comparative experimental biology.

[36]  G. Acland,et al.  Comparative analysis and expression of CLUL1, a cone photoreceptor-specific gene. , 2003, Investigative ophthalmology & visual science.

[37]  Stuart K. Kim,et al.  Gene expression analysis in a transgenic Caenorhabditis elegans Alzheimer’s disease model , 2003, Neurobiology of Aging.

[38]  K. Ray,et al.  Molecular cloning, characterization and expression of a novel retinal clusterin-like protein cDNA. , 2000, Gene.

[39]  T. Shows,et al.  Characterization of a novel tumor necrosis factor-alpha-induced endothelial primary response gene. , 1992, The Journal of biological chemistry.

[40]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[41]  B. Thisse,et al.  High-resolution in situ hybridization to whole-mount zebrafish embryos , 2007, Nature Protocols.

[42]  M. Westerfield The zebrafish book : a guide for the laboratory use of zebrafish (Danio rerio) , 1995 .