Received date: 2022-08-01
Revised date: 2022-09-20
Online published: 2022-10-03
Supported by
the Training Program of the Major Research Plan of the National Natural Science Foundation of China(91857110);the National Key Research and Development Program of China(2018YFA0800403);the National Key Research and Development Program of China(2016YFC1305303);the National Natural Science Foundation of China(81670740);the National Natural Science Fund for Excellent Young Scholars of China(81722012);the Zhejiang Provincial Natural Science Foundation of China(LZ21H070001);the Innovative Institute of Basic Medical Sciences ofZhejiang University, and the Fundamental Research Funds for the Central Universities, the Construction Fund of Medical Key Disciplinesof Hangzhou(OO20200055);K.C. Wong Education Foundation
Cleavage under target and tagment (CUT&Tag) is a technology that utilizes the fusion protein of Tn5 transposase and protein A/G which can guide Tn5 enzyme to the antibody bound to target protein and cleave the chromatin regions adjacent to target protein. Chromatin libraries are then tagged and sequenced by the high-throughput sequencing to obtain chromatin information at specific sites or protein binding locations. CUT&Tag technology plays an important role in the research of DNA and protein interactions. It can be used to understand the modifications of histone and the bindings of transcription factors. Compared with the traditional chromatin immunoprecipitation-sequencing (ChIP-seq) technology, the CUT&Tag has the strengths of high signal-to-noise ratio, good repeatability, short experimental period, and low cell input. It shows great advantages in early embryonic development, stem cells, cancer, epigenetics and other research fields. In this article, we described the protocol of CUT&Tag for metabolic tissue cells (mouse primary islet cells), to provide an epigenetic method for studying metabolic cells.
Key words: CUT&Tag; histone modification; protocol; metabolic cell
Ziyin Zhang, Yanping Zhou, Zhuo-Xian Meng . The protocol of CUT&Tag for metabolic tissue cells[J]. Hereditas(Beijing), 2022 , 44(10) : 958 -966 . DOI: 10.16288/j.yczz.22-253
| [1] | Kidder BL, Hu GQ, Zhao KJ. ChIP-Seq: technical considerations for obtaining high-quality data. Nat Immunol, 2011, 12(10): 918-922. |
| [2] | Kaya-Okur HS, Wu SJ, Codomo CA, Pledger ES, Bryson TD, Henikoff JG, Ahmad K, Henikoff S. CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nat Commun, 2019, 10(1): 1930. |
| [3] | Henikoff S, Henikoff JG, Kaya-Okur HS, Ahmad K. Efficient chromatin accessibility mapping in situ by nucleosome-tethered tagmentation. eLife, 2020, 9: e63274. |
| [4] | Kaya-Okur HS, Janssens DH, Henikoff JG, Ahmad K, Henikoff S. Efficient low-cost chromatin profiling with CUT&Tag. Nat Protoc, 2020, 15(10): 3264-3283. |
| [5] | Wei Y, Li K, Lu DR, Zhu HX. Optimization of CUT&Tag product recovery and library construction method. Hereditas (Beijing), 2021, 43(4): 362-374. |
| [5] | 韦晔, 李科, 卢大儒, 朱化星. CUT&Tag产物回收和建库方法的优化. 遗传, 2021, 43(4): 362-374. |
| [6] | Taylor BL, Liu FF, Sander M. Nkx6.1 is essential for maintaining the functional state of pancreatic beta cells. Cell Rep, 2013, 4(6): 1262-1275. |
| [7] | Qiao JT, Zhang ZY, Ji SH, Liu TL, Zhang XN, Huang YM, Feng WL, Wang KL, Wang JY, Wang SS, Meng ZX, Liu M. A distinct role of STING in regulating glucose homeostasis through insulin sensitivity and insulin secretion. Proc Natl Acad Sci USA, 2022, 119(7): e2101848119. |
| [8] | Wang RR, Qiu XY, Pan R, Fu HX, Zhang ZY, Wang QT, Chen HD, Wu QQ, Pan XW, Zhou YP, Shan PF, Wang SS, Guo GJ, Zheng M, Zhu LY, Meng ZX. Dietary intervention preserves β cell function in mice through CTCF-mediated transcriptional reprogramming. J Exp Med, 2022, 219(7): e20211779. |
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