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Disconnected interacting protein 2 homolog A (DIP2A) is highly expressed in nervous system and respiratory system of developing embryos. However, genes regulated by Dip2a in developing brain and lung have not been systematically studied. Transcriptome of brain and lung in embryonic 19.5 day (E19.5) were compared between wild type and Dip2a -/- mice. An average of 50 million reads per sample was mapped to the reference sequence. A total of 214 DEGs were detected in brain (82 up and 132 down) and 1900 DEGs in lung (1259 up and 641 down). GO enrichment analysis indicated that DEGs in both Brain and Lung were mainly enriched in biological processes ‘DNA-templated transcription and Transcription from RNA polymerase II promoter’, ‘multicellular organism development’, ‘cell differentiation’ and ‘apoptotic process’. In addition, COG classification showed that both were mostly involved in ‘Replication, Recombination, and Repair’, ‘Signal transduction and mechanism’, ‘Translation, Ribosomal structure and Biogenesis’ and ‘Transcription’. KEGG enrichment analysis showed that brain was mainly enriched in ‘Thyroid cancer’ pathway whereas lung in ‘Complement and Coagulation Cascades’ pathway. Transcription factor (TF) annotation analysis identified Zinc finger domain containing (ZF) proteins were mostly regulated in lung and brain. Interestingly, study identified genes Skor2 , Gpr3711 , Runx1 , Erbb3 , Frmd7 , Fut10 , Sox11 , Hapln1 , Tfap2c and Plxnb3 from brain that play important roles in neuronal cell maturation, differentiation, and survival; genes Hoxa5 , Eya1 , Errfi1 , Sox11 , Shh , Igf1 , Ccbe1 , Crh , Fgf9 , Lama5 , Pdgfra , Ptn , Rbp4 and Wnt7a from lung are important in lung development. Expression levels of the candidate genes were validated by qRT-PCR. Genome wide transcriptional analysis using wild type and Dip2a knockout mice in brain and lung at embryonic day 19.5 (E19.5) provided a genetic basis of molecular function of these genes.

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14Disconnected interacting 2 homolog A (DIP2A) gene is highly 15 expressed in nervous system and respiratory system of developing 16 embryos. However, genes regulated by Dip2a in developing brain and 17 lung have not been systemically studied. Transcriptome of brain and lung 18 in embryonic 19.5 day (E19.5) were compared between wild type and 19 Dip2a -/mice. Total RNAs were extracted from brain and lung of E19.5 20 embryos for RNA-Seq. Clean reads were mapped to mouse reference 21 sequence (mm9) using Tophat and assembled into transcripts by 22 Cufflinks. Edge R and DESeq were applied to identify differentially 2 23 expressed genes (DEGs) and annotated under GO, COG, KEGG and TF. 24An average of 50 million reads per sample was mapped to the reference 25 sequence. A total of 214 DEGs were detected in brain (82 up and 132 26 down) and 1900 DEGs in lung (1259 up and 641 down). GO enrichment 27 analysis indicated that DEGs in both Brain and Lung were mainly 28 enriched in biological processes 'DNA-templated transcription and 29 Transcription from RNA polymerase II promoter', 'multicellular 30 organism development', 'cell differentiation' and 'apoptotic process'. In 31 addition, COG classification showed that both were mostly involved in 32 'Replication, Recombination and Repair', 'Signal transduction and 33 mechanism', 'Translation, Ribosomal structure and Biogenesis' and 34 'Transcription'. KEGG enrichment analysis showed that brain was 35 mainly enriched in 'Thryoid cancer' pathway whereas lung in 36 'Complement and Coagulation Cascades' pathway. Transcription factor 37 (TF) annotation analysis identified Zinc finger domain containing (ZF) 38 proteins were mostly regulated in lung and brain. Interestingly, study 39 identified genes Skor2, Gpr3711, Runx1, Erbb3, Frmd7, Fut10, Sox11, 40 Hapln1, Tfap2c and Plxnb3 from brain that play important roles in 41 neuronal cell maturation, differentiation and survival; genes Hoxa5, 42 Eya1, Ctsh, Erff1, Lama1, Lama2, Rspo2, Sox11, Spry4, Shh, Igf1 and 43 Wnt7a from lung are important in lung development and morphogenesis. 44 Expression levels of the candidate genes were validated by qRT-PCR. 3 45 Genome wide transcriptional analysis using wild type and Dip2a 46 knockout mice in brain and lung at embryonic day 19.5 (E19.5) provided 47 [7-9]. These all evidence strongly supports the role of Dip2a gene in both 65 vertebrate and invertebrate nervous system development. However, 66 which biological process or molecular function is regulated by Dip2a 4 67 gene during embryonic brain development is not known. 68 In order to systematically understand expression pattern and 69 physiological role of Dip2a gene, Dip2a-LacZ knockin and Dip2a 65kb 70 knockout (Dip2a -/-) mouse models were generated using CRISPR/Cas9 71 system [10]. Dip2a expression via endogenous expression of 72 β-Galactosidase gene (LacZ) have shown that Dip2a is highly expressed 73 in brain neurons, retinal ganglia cell, reproductive, vascular and Lung 74 tissue in adult and ectodermal tissue in developing embryos [11]...