[No authors listed]
The MSM/Ms strain is derived from the Japanese wild mouse Mus musculus molossinus and displays characteristics not observed in common laboratory strains. Functional genomic analyses using genetically engineered MSM/Ms mice will reveal novel phenotypes and gene functions/interactions. We previously reported the establishment of a germline-competent embryonic stem (ES) cell line, Mol/MSM-1, from the MSM/Ms strain. To analyze its usefulness for insertional mutagenesis, we performed gene-trapping using these cells. In the present study, we compared the gene-trap events between Mol/MSM-1 and a conventional ES cell line, KTPU8, derived from the F1 progeny of a C57BL/6 Ã CBA cross. We introduced a promoter-trap vector carrying the promoterless β-galactosidase/neomycin-resistance fusion gene into Mol/MSM-1 and KTPU8 cells, isolated clones, and identified the trapped genes by rapid amplification of cDNA 5'-ends (5'-RACE), inverse PCR, or plasmid rescue. Unexpectedly, the success rate of 5'-RACE in Mol/MSM trap clones was 47 %, lower than the 87 % observed in KTPU8 clones. Genomic analysis of the 5'-RACE-failed clones revealed that most had trapped ribosomal RNA gene regions. The percentage of ribosomal RNA region trap clones was 41 % in Mol/MSM-1 cells, but less than 10 % in KTPU8 cells. However, within the Mol/MSM-1 5'-RACE-successful clones, the trapping frequency of annotated genes, the chromosomal distribution of vector insertions, the frequency of integration into an intron around the start codon-containing exon, and the functional spectrum of trapped genes were comparable to those in KTPU8 cells. By selecting 5'-RACE-successful clones, it is possible to perform gene-trapping efficiently using Mol/MSM-1 ES cells and promoter-trap vectors.
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Rex2, Cnot6, Foxp1, Mta1, Pop5, Bmpr1a, Dnajc5, Ctbp2, Celf1, Mark2, Sfxn1, Gli2, Klf12, Kpna2, Alad, Nedd4, Pkig, Plcb4, Pvt1, Rbmxl1, Rbpms, Rbpj, Ralbp1, Rnf2, Stmn2, Camkk2, Etl4, Hmgcs1, Daam1, Tnrc6b, Tead1, Tmem11, Itpk1, Arl15, Hmbox1, Traf3, Tsix, Slc25a51, Tnrc6a, Tmem161a, Usp32, Abcc4, Pacsin2, Zfp407, G3bp1, Txnl4a, Sox2ot, Mreg, Snhg3, Zdhhc18, Txnrd1, Carhsp1, Rev1, Vapb, Chst12, Rpl23, Cyb5b, Psmd12, Rnmt, Setd8, Rps13, 1110002J07Rik, 2410006H16Rik, Qrich1, Wdfy1, Ten1, Mier2, Mutyh, Ist1, Fryl, Rab10os, Fblim1, Tti1, Nln, Msi2, Csrnp3, Sh3bp4
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