一、 DAP-seq技术简介

DAP-seq（DNA亲和纯化测序）技术服务——高通量检测转录因子或DNA结合蛋白在基因组上的结合位点。

在功能基因组学和表观遗传学研究中，转录因子结合位点（TFBS）的发掘一直是研究热点。传统的ChIP-seq（染色质免疫共沉淀测序）方法，在抗体质量很好的情况下能够有效检测到TFBS。然而，好的抗体可遇不可求，这限制了ChIP-seq更广泛的应用。

2016年，O'Malley RC等人在Cell上发表了使用DAP-seq技术，快速绘制转录因子调控靶向DNA区域图谱的文章。2017年，Bartlett A等人在Nature Protocol上发表了 DAP-seq的实验方法。 DAP-seq技术的出现，使TFBS 的研究不再局限于物种，不再受抗体质量的限制，为生命科学和医学领域转录因子的研究提供了新的有效工具。

二、 ChIP-seq和DAP-seq技术对比

三、DAP-seq技术流程：

蓝景科信已发表文章列表：

Zhang SL, Wang L, Yao J, Wu N, Ahmad B, Nocker S, Wu JY, Abudureheman R, Li Z, Wang XP. Control of ovule development in Vitis vinifera by VvMADS28 and interacting genes. Horticulture Research. 2023. doi: 10.1093/hr/uhad070. (IF=7.291) 

Wang L, Tian T, Liang J, Li R, Xin X, Qi Y, Zhou Y, Fan Q, Ning G, Becana M, Duanmu D. A transcription factor of the NAC family regulates nitrate-induced legume nodule senescence. New Phytol. 2023 Mar 22. doi: 10.1111/nph.18896. (IF=10.323)

Sun Y, Han Y, Sheng K, Yang P, Cao Y, Li H, Zhu QH, Chen J, Zhu S, Zhao T. Single-cell transcriptomic analysis reveals the developmental trajectory and transcriptional regulatory networks of pigment glands in Gossypium bickii. Mol Plant. 2023. doi: 10.1016/j.molp.2023.02.005. (IF=21.949) 

Liu Y, Liu Q, Li X, Zhang Z, Ai S, Liu C, Ma F, Li C. MdERF114 enhances the resistance of apple roots to Fusarium solani by regulating the transcription of MdPRX63. Plant Physiol. 2023. doi: 10.1093/plphys/kiad057. (IF=8.005) 

Liu YN, Wu FY, Tian RY, Shi YX, Xu ZQ, Liu JY, Huang J, Xue FF, Liu BY, Liu GQ. The bHLH-zip transcription factor SREBP regulates triterpenoid and lipid metabolisms in the medicinal fungus Ganoderma lingzhi. Commun Biol. 2023. doi: 10.1038/s42003-022-04154-6. (IF=6.548)

Liu L, Chen G, Li S, Gu Y, Lu L, Qanmber G, Mendu V, Liu Z, Li F, Yang Z. A brassinosteroid transcriptional regulatory network participates in regulating fiber elongation in cotton. Plant Physiol. 2022. doi: 10.1093/plphys/kiac590. (IF=8.005)

Li M, Hou L, Zhang C, Yang W, Liu X, Zhao H, Pang X, Li Y. Genome-Wide Identification of Direct Targets of ZjVND7 Reveals the Putative Roles of Whole-Genome Duplication in Sour Jujube in Regulating Xylem Vessel Differentiation and Drought Tolerance. Front Plant Sci. 2022 Feb 4;13:829765. doi: 10.3389/fpls.2022.829765. (IF=6.627)

Bi Y, Wang H, Yuan X, Yan Y, Li D, Song F. The NAC transcription factor ONAC083 negatively regulates rice immunity against Magnaporthe oryzae by directly activating transcription of the RING-H2 gene OsRFPH2-6. J Integr Plant Biol. 2022. doi: 10.1111/jipb.13399. (IF=9.106)

Guo X, Yu X, Xu Z, Zhao P, Zou L, Li W, Geng M, Zhang P, Peng M, Ruan M. CC-type glutaredoxin, MeGRXC3, associates with catalases and negatively regulates drought tolerance in cassava (Manihot esculenta Crantz). Plant Biotechnol J. 2022. doi: 10.1111/pbi.13920. (IF=13.263)

Chai Z, Fang J, Huang C, Huang R, Tan X, Chen B, Yao W, Zhang M. A novel transcription factor, ScAIL1, modulates plant defense responses by targeting DELLA and regulating gibberellin and jasmonic acid signaling in sugarcane. J Exp Bot. 2022. 73: 6727-6743. doi: 10.1093/jxb/erac339. (IF=7.298)

Li R, Zheng W, Yang R, Hu Q, Ma L, Zhang H. OsSGT1 promotes melatonin-ameliorated seed tolerance to chromium stress by affecting the OsABI5-OsAPX1 transcriptional module in rice. Plant J. 2022. 112: 151-171. doi: 10.1111/tpj.15937. (IF=5.726)

Li Q, Zhou L, Chen Y, Xiao N, Zhang D, Zhang M, Wang W, Zhang C, Zhang A, Li H, Chen J, Gao Y. Phytochrome interacting factor regulates stomatal aperture by coordinating red light and abscisic acid. Plant Cell. 2022. 34: 4293-4312. doi: 10.1093/plcell/koac244. (IF=12.085)

Luo M, Lu B, Shi Y, Zhao Y, Wei Z, Zhang C, Wang Y, Liu H, Shi Y, Yang J, Song W, Lu X, Fan Y, Xu L, Wang R, Zhao J. A newly characterized allele of ZmR1 increases anthocyanin content in whole maize plant and the regulation mechanism of different ZmR1 alleles. Theor Appl Genet. 2022. 135: 3039-3055. doi: 10.1007/s00122-022-04166-0. (IF=5.574)

Wei H, Xu H, Su C, Wang X, Wang L. Rice CIRCADIAN CLOCK ASSOCIATED 1 transcriptionally regulates ABA signaling to confer multiple abiotic stress tolerance. Plant Physiol. 2022. 190: 1057-1073. doi: 10.1093/plphys/kiac196. (IF=8.005)

Tang N, Cao Z, Yang C, Ran D, Wu P, Gao H, He N, Liu G, Chen Z. A R2R3-MYB transcriptional activator LmMYB15 regulates chlorogenic acid biosynthesis and phenylpropanoid metabolism in Lonicera macranthoides. Plant Sci. 2021. 308: 110924. doi: 10.1016/j.plantsci.2021.110924. (IF=5.363)

Liang S, Gao X, Wang Y, Zhang H, Yin K, Chen S, Zhang M, Zhao R. Phytochrome-interacting factors regulate seedling growth through ABA signaling. Biochem Biophys Res Commun. 2020. 526: 1100-1105. doi: 10.1016/j.bbrc.2020.04.011. (IF=3.322)

Yao J, Shen Z, Zhang Y, Wu X, Wang J, Sa G, Zhang Y, Zhang H, Deng C, Liu J, Hou S, Zhang Y, Zhang Y, Zhao N, Deng S, Lin S, Zhao R, Chen S. Populus euphratica WRKY1 binds the promoter of H+-ATPase gene to enhance gene expression and salt tolerance. J Exp Bot. 2020. 71: 1527-1539. doi: 10.1093/jxb/erz493. (IF=5.36)

四、DAP-seq实验流程

五、DAP-seq生信分析内容

1.对原始数据进行去除接头、污染序列及低质量 reads 的处理

2.数据产出统计

3.参考序列比对分析 

4.测序reads富集区域扫描（peak calling）

5.Peak长度分布统计

6.Peak在基因功能元件上的分布统计

7.Peak序列模式发掘（motif search）

8.已知motif注释

9.Peak相关基因鉴定

10.Peak相关基因的GO和KEGG富集分析

11.测序数据的差异分析（>=2个样本）

12.测序数据的可视化分析