دورية أكاديمية
TALEN-Mediated FLAG-Tagging of Endogenous Histone Methyltransferase DOT1L.
| العنوان: | TALEN-Mediated FLAG-Tagging of Endogenous Histone Methyltransferase DOT1L. |
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| المؤلفون: | An C; Guang'an Men Hospital, China Academy of Chinese Medical Sciences, Beijing, China.; Laboratory of Human Environmental Epigenomes, Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA., Zhu G; Laboratory of Human Environmental Epigenomes, Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA., Martos SN; Laboratory of Human Environmental Epigenomes, Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA., Feng X; Guang'an Men Hospital, China Academy of Chinese Medical Sciences, Beijing, China., Zhang H; School of Life Sciences, Hubei University, Wuhan, China., Jia Y; GENEWIZ Suzhou, Suzhou, China., Wang Z; Laboratory of Human Environmental Epigenomes, Department of Environmental Health Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, USA.; School of Life Sciences, Hubei University, Wuhan, China.; Fenxian Central Hospital, Shanghai, China.; Department of Oncology and Sidney Kimmel Comprehensive Cancer Center, School of Medicine, Johns Hopkins University, Baltimore, MD, USA. |
| المصدر: | Advances in bioscience and biotechnology (Print) [Adv Biosci Biotechnol] 2017 Sep; Vol. 8 (9), pp. 311-323. Date of Electronic Publication: 2017 Sep 22. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Scientific Research Publishing, Inc Country of Publication: United States NLM ID: 101553812 Publication Model: Print-Electronic Cited Medium: Print ISSN: 2156-8456 (Print) NLM ISO Abbreviation: Adv Biosci Biotechnol Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Irvine, CA : Scientific Research Publishing, Inc. |
| مستخلص: | Histone modification including H3 lysine 79 methylation (H3K79me) plays a key role during gene transcription and DNA damage repair. DOT1L, the sole methyltransferase for three states of H3K79me, is implicated in leukemia, co-lorectal cancer, and dilated cardiomyopathy. However, understanding of DOT1L and H3K79me in these pathways and disease pathogenesis has been limited due to the difficulty of working with DOT1L protein. For instance, locus-specific or genome-wide binding sites of DOT1L revealed by chromatin immunoprecipitation (ChIP)-based methods are necessary for inferring its functions, but high-quality ChIP-grade antibodies are currently not available. Herein we have developed a knock-in approach to tag endogenous DOT1L with 3 × Flag at its C-terminal domain to follow functional analyses. The knock-in was facilitated by using TALENs to induce a targeted double-strand break at the endogenous DOTIL to stimulate local homologous recombination at that site. The single cell colonies with successful knock-in were isolated and verified by different methods. We also demonstrated that tagged DOT1L maintains its normal function in terms of methylation and that the engineered cells would be very useful for further studies. Competing Interests: Conflict of Interest Z.W serves as a consultant for CUSABIO (CusAb) Company, Wuhan, China. |
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| معلومات مُعتمدة: | R01 ES025761 United States ES NIEHS NIH HHS; T32 ES007141 United States ES NIEHS NIH HHS; U01 ES026721 United States ES NIEHS NIH HHS |
| فهرسة مساهمة: | Keywords: DOT1L; Flag; Knock-In; TALEN |
| تواريخ الأحداث: | Date Created: 20180526 Latest Revision: 20240610 |
| رمز التحديث: | 20240610 |
| مُعرف محوري في PubMed: | PMC5963693 |
| DOI: | 10.4236/abb.2017.89023 |
| PMID: | 29796335 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2156-8456 |
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| DOI: | 10.4236/abb.2017.89023 |