Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9
Pancreatic cancer is one of the most painful and aggressive types of cancer with high lethality rate. Early diagnosis of the disease is limited and antitumor therapy has low efficacy, because pancreatic cancer shows multiple drug resistance. It is still unclear how pancreatic tumor cells become resi...
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| Format: | Article |
| Language: | English |
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Kazan Federal University
2018-09-01
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| Series: | Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki |
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| Online Access: | https://kpfu.ru/obtaining-a-pancreatic-cancer-cell-line-stably_382366.html |
| _version_ | 1797810759553515520 |
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| author | A.K. Nurgalieva V.S. Skripova L.F. Minigulova R.G. Kiyamova |
| author_facet | A.K. Nurgalieva V.S. Skripova L.F. Minigulova R.G. Kiyamova |
| author_sort | A.K. Nurgalieva |
| collection | DOAJ |
| description | Pancreatic cancer is one of the most painful and aggressive types of cancer with high lethality rate. Early diagnosis of the disease is limited and antitumor therapy has low efficacy, because pancreatic cancer shows multiple drug resistance. It is still unclear how pancreatic tumor cells become resistant to chemotherapy and understanding of the resistance mechanisms and identification of their regulating genes is necessary to develop a new and more efficient pancreatic cancer therapy. One of the methods to identify such genes is CRISPR/Cas9 technology (clustered regularly interspaced short palindromic repeats) where Cas9 endonuclease is used to make double-strand DNA breaks in target genes.
The aim of this work is modification of the pancreatic cancer cell line MIA PaCa-2 to express a doxycycline-inducible Cas9 endonuclease gene. Several plasmid vectors have been used to create a lentivirus containing the Cas9 endonuclease gene. The virus has been used for transduction of tumor cells to get doxycycline-inducible Cas9 expressing clones. Cas9 endonuclease expression has been detected by the western blot analysis with antibodies to FLAG epitope.
Two tumor clones of the MIA Pa-Ca2/Cas9 cell line expressing Cas9 endonuclease have been generated. These cells can be used further to search genes related to regulation of the susceptibility to antitumor drugs. |
| first_indexed | 2024-03-13T07:13:36Z |
| format | Article |
| id | doaj.art-136d82416f4b4a8880ba2db13e589cd1 |
| institution | Directory Open Access Journal |
| issn | 2542-064X 2500-218X |
| language | English |
| last_indexed | 2024-03-13T07:13:36Z |
| publishDate | 2018-09-01 |
| publisher | Kazan Federal University |
| record_format | Article |
| series | Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki |
| spelling | doaj.art-136d82416f4b4a8880ba2db13e589cd12023-06-05T14:24:55ZengKazan Federal UniversityUčënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki2542-064X2500-218X2018-09-011603386394Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9A.K. Nurgalieva0V.S. Skripova1L.F. Minigulova2R.G. Kiyamova3Kazan Federal University, Kazan, 420008 RussiaKazan Federal University, Kazan, 420008 RussiaKazan Federal University, Kazan, 420008 RussiaKazan Federal University, Kazan, 420008 RussiaPancreatic cancer is one of the most painful and aggressive types of cancer with high lethality rate. Early diagnosis of the disease is limited and antitumor therapy has low efficacy, because pancreatic cancer shows multiple drug resistance. It is still unclear how pancreatic tumor cells become resistant to chemotherapy and understanding of the resistance mechanisms and identification of their regulating genes is necessary to develop a new and more efficient pancreatic cancer therapy. One of the methods to identify such genes is CRISPR/Cas9 technology (clustered regularly interspaced short palindromic repeats) where Cas9 endonuclease is used to make double-strand DNA breaks in target genes. The aim of this work is modification of the pancreatic cancer cell line MIA PaCa-2 to express a doxycycline-inducible Cas9 endonuclease gene. Several plasmid vectors have been used to create a lentivirus containing the Cas9 endonuclease gene. The virus has been used for transduction of tumor cells to get doxycycline-inducible Cas9 expressing clones. Cas9 endonuclease expression has been detected by the western blot analysis with antibodies to FLAG epitope. Two tumor clones of the MIA Pa-Ca2/Cas9 cell line expressing Cas9 endonuclease have been generated. These cells can be used further to search genes related to regulation of the susceptibility to antitumor drugs.https://kpfu.ru/obtaining-a-pancreatic-cancer-cell-line-stably_382366.htmlpancreatic cancercrispr/cas9cas9 endonucleasemia paca-2 |
| spellingShingle | A.K. Nurgalieva V.S. Skripova L.F. Minigulova R.G. Kiyamova Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 Učënye Zapiski Kazanskogo Universiteta. Seriâ Estestvennye Nauki pancreatic cancer crispr/cas9 cas9 endonuclease mia paca-2 |
| title | Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 |
| title_full | Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 |
| title_fullStr | Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 |
| title_full_unstemmed | Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 |
| title_short | Obtaining a pancreatic cancer cell line stably expressing doxycycline-dependent endonuclease Cas9 |
| title_sort | obtaining a pancreatic cancer cell line stably expressing doxycycline dependent endonuclease cas9 |
| topic | pancreatic cancer crispr/cas9 cas9 endonuclease mia paca-2 |
| url | https://kpfu.ru/obtaining-a-pancreatic-cancer-cell-line-stably_382366.html |
| work_keys_str_mv | AT aknurgalieva obtainingapancreaticcancercelllinestablyexpressingdoxycyclinedependentendonucleasecas9 AT vsskripova obtainingapancreaticcancercelllinestablyexpressingdoxycyclinedependentendonucleasecas9 AT lfminigulova obtainingapancreaticcancercelllinestablyexpressingdoxycyclinedependentendonucleasecas9 AT rgkiyamova obtainingapancreaticcancercelllinestablyexpressingdoxycyclinedependentendonucleasecas9 |