CRISPR/Cas9-mediated knockout of c-REL in HeLa cells results in profound defects of the cell cycle
Publication Date
August 02, 2017
Authors
Carsten Slotta, Thomas Schlüter, Lucia M. Ruiz Perera, Hussamadin M. Kadhim, et al
Volume
12
Issue
8
Pages
e0182373
DOI
https://dx.plos.org/10.1371/journal.pone.0182373
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0182373
Scopus
85026649697
Mendeley
http://www.mendeley.com/research/crisprcas9mediated-knockout-crel-hela-cells-results-profound-defects-cell-cycle
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Mendeley | Further Information

{"title"=>"CRISPR/Cas9-mediated knockout of c-REL in HeLa cells results in profound defects of the cell cycle", "type"=>"journal", "authors"=>[{"first_name"=>"Carsten", "last_name"=>"Slotta", "scopus_author_id"=>"55935791700"}, {"first_name"=>"Thomas", "last_name"=>"Schlüter", "scopus_author_id"=>"6603863936"}, {"first_name"=>"Lucia M.", "last_name"=>"Ruiz-Perera", "scopus_author_id"=>"56008726000"}, {"first_name"=>"Hussamadin M.", "last_name"=>"Kadhim", "scopus_author_id"=>"57195259876"}, {"first_name"=>"Tobias", "last_name"=>"Tertel", "scopus_author_id"=>"57195265086"}, {"first_name"=>"Elena", "last_name"=>"Henkel", "scopus_author_id"=>"57195261549"}, {"first_name"=>"Wolfgang", "last_name"=>"Hübner", "scopus_author_id"=>"7004332955"}, {"first_name"=>"Johannes F.W.", "last_name"=>"Greiner", "scopus_author_id"=>"55068100500"}, {"first_name"=>"Thomas", "last_name"=>"Huser", "scopus_author_id"=>"7004227222"}, {"first_name"=>"Barbara", "last_name"=>"Kaltschmidt", "scopus_author_id"=>"6701680723"}, {"first_name"=>"Christian", "last_name"=>"Kaltschmidt", "scopus_author_id"=>"7003915652"}], "year"=>2017, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"617606380", "doi"=>"10.1371/journal.pone.0182373", "scopus"=>"2-s2.0-85026649697", "pmid"=>"28767691", "sgr"=>"85026649697", "issn"=>"19326203", "isbn"=>"1111111111"}, "id"=>"e0817f4f-2766-3b32-8c5d-95a9b9c580f5", "abstract"=>"Cervical cancer is the fourth common cancer in women resulting worldwide in 266,000 deaths per year. Belonging to the carcinomas, new insights into cervical cancer biology may also have great implications for finding new treatment strategies for other kinds of epithelial cancers. Although the transcription factor NF-κB is known as a key player in tumor formation, the relevance of its particular subunits is still underestimated. Here, we applied CRISPR/Cas9n-mediated genome editing to successfully knockout the NF-κB subunit c-REL in HeLa Kyoto cells as a model system for cervical cancers. We successfully generated a homozygous deletion in the c-REL gene, which we validated using sequencing, qPCR, immunocytochemistry, western blot analysis, EMSA and analysis of off-target effects. On the functional level, we observed the deletion of c-REL to result in a significantly decreased cell proliferation in comparison to wildtype (wt) without affecting apoptosis. The impaired proliferative behavior of c-REL-/- cells was accompanied by a strongly decreased amount of the H2B protein as well as a significant delay in the prometaphase of mitosis compared to c-REL+/+ HeLa Kyoto cells. c-REL-/- cells further showed significantly decreased expression levels of c-REL target genes in comparison to wt. In accordance to our proliferation data, we observed the c-REL knockout to result in a significantly increased resistance against the chemotherapeutic agents 5-Fluoro-2’-deoxyuridine (5-FUDR) and cisplatin. In summary, our findings emphasize the importance of c-REL signaling in a cellular model of cervical cancer with direct clinical implications for the development of new treatment strategies.", "link"=>"http://www.mendeley.com/research/crisprcas9mediated-knockout-crel-hela-cells-results-profound-defects-cell-cycle", "reader_count"=>10, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>4, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Student > Doctoral Student"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>4, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>4}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>1}}, "group_count"=>0}

Scopus | Further Information

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