Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation
The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors...
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| Format: | Article |
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MDPI AG
2018-06-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/23/6/1360 |
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| author | Hyun-Ji Choi Han-Byul Lee Sunyoung Jung Hyun-Kyu Park Woori Jo Sung-Min Cho Woo-Jin Kim Woo-Chan Son |
| author_facet | Hyun-Ji Choi Han-Byul Lee Sunyoung Jung Hyun-Kyu Park Woori Jo Sung-Min Cho Woo-Jin Kim Woo-Chan Son |
| author_sort | Hyun-Ji Choi |
| collection | DOAJ |
| description | The Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors in the prostate. In the present study, electroporation was used to enhance the transfection efficiency of the SB transposon. To generate tumors, three constructs (a c-Myc expression cassette, a HRAS (HRas proto-oncogene, GTPase) expression cassette and a shRNA against p53) contained within the SB transposon plasmids were directly injected into the prostate. Electroporation was conducted on the injection site after the injection of the DNA plasmid. Following the tumorigenesis, the tumors were monitored by animal PET imaging and identified by gross observation. After this, the tumors were characterized by using histological and immunohistochemical techniques. The expression of the targeted genes was analyzed by Real-Time qRT-PCR. All mice subjected to the injection were found to have prostate tumors, which was supported by PSA immunohistochemistry. To our knowledge, this is the first demonstration of tumor induction in the mouse prostate using the electroporation-enhanced SB transposon system in combination with c-Myc, HRAS and p53. This model serves as a valuable resource for the future development of SB-induced mouse models of cancer. |
| first_indexed | 2024-12-11T22:35:41Z |
| format | Article |
| id | doaj.art-fc456e60e2314b8ea007390b4206dac7 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-11T22:35:41Z |
| publishDate | 2018-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-fc456e60e2314b8ea007390b4206dac72022-12-22T00:47:58ZengMDPI AGMolecules1420-30492018-06-01236136010.3390/molecules23061360molecules23061360Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and ElectroporationHyun-Ji Choi0Han-Byul Lee1Sunyoung Jung2Hyun-Kyu Park3Woori Jo4Sung-Min Cho5Woo-Jin Kim6Woo-Chan Son7Asan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaDepartment of Pathology, University of Ulsan College of Medicine, Songpa-gu, 05505 Seoul, KoreaAsan Institute for Life Sciences, Asan Medical Center, Songpa-gu, 05505 Seoul, KoreaThe Sleeping Beauty (SB) transposon system is non-viral and uses insertional mutagenesis, resulting in the permanent expression of transferred genes. Although the SB transposon is a useful method for establishing a mouse tumor model, there has been difficulty in using this method to generate tumors in the prostate. In the present study, electroporation was used to enhance the transfection efficiency of the SB transposon. To generate tumors, three constructs (a c-Myc expression cassette, a HRAS (HRas proto-oncogene, GTPase) expression cassette and a shRNA against p53) contained within the SB transposon plasmids were directly injected into the prostate. Electroporation was conducted on the injection site after the injection of the DNA plasmid. Following the tumorigenesis, the tumors were monitored by animal PET imaging and identified by gross observation. After this, the tumors were characterized by using histological and immunohistochemical techniques. The expression of the targeted genes was analyzed by Real-Time qRT-PCR. All mice subjected to the injection were found to have prostate tumors, which was supported by PSA immunohistochemistry. To our knowledge, this is the first demonstration of tumor induction in the mouse prostate using the electroporation-enhanced SB transposon system in combination with c-Myc, HRAS and p53. This model serves as a valuable resource for the future development of SB-induced mouse models of cancer.http://www.mdpi.com/1420-3049/23/6/1360electroporationinsertional mutagenesisanimal modelsprostatic neoplasmsSleeping Beauty transposase |
| spellingShingle | Hyun-Ji Choi Han-Byul Lee Sunyoung Jung Hyun-Kyu Park Woori Jo Sung-Min Cho Woo-Jin Kim Woo-Chan Son Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation Molecules electroporation insertional mutagenesis animal models prostatic neoplasms Sleeping Beauty transposase |
| title | Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation |
| title_full | Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation |
| title_fullStr | Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation |
| title_full_unstemmed | Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation |
| title_short | Development of a Mouse Model of Prostate Cancer Using the Sleeping Beauty Transposon and Electroporation |
| title_sort | development of a mouse model of prostate cancer using the sleeping beauty transposon and electroporation |
| topic | electroporation insertional mutagenesis animal models prostatic neoplasms Sleeping Beauty transposase |
| url | http://www.mdpi.com/1420-3049/23/6/1360 |
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