NanoTag - an IgG-free method for mapping DNA-protein interactions
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Date
2024-07-12
Publication Type
Working Paper
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yes
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Abstract
Genome-wide profiling of DNA-protein interactions in cells can provide important information 20 about mechanisms of gene regulation. Most current methods for genome-wide profiling of 21 DNA-bound proteins, such as ChIP-seq and CUT&Tag, use conventional IgG antibodies to 22 bind target protein(s), which limits their applicability to targets for which high affinity and high 23 specificity antibodies are available. Here we describe NanoTag, a novel method derived from 24 CUT&Tag that is IgG-free and uses a nanobody to profile DNA-protein interactions. NanoTag 25 is based on an anti-GFP nanobody-Tn5 transposase fusion that allows mapping GFP-tagged 26 proteins associated with chromatin in a fast and cost-effective manner. We demonstrate the 27 utility of NanoTag by profiling the histone mark H3K4me3 via its binding partner TATA box 28 binding protein-associated factor 3 (TAF3) and the transcription factors Nanog and CTCF in 29 mouse embryonic stem cells expressing GFP-tagged targets. For the targets examined, 30 NanoTag data shows high correlation to CUT&Tag data and displays a similarly high signal 31 to-noise ratio. Overall, NanoTag provides a flexible, IgG-free and cost-effective method to 32 generate high resolution DNA-binding profiles in cells or tissues.
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published
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Publisher
Cold Spring Harbor Laboratory
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Edition / version
v1
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Software
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Subject
Chromatin; DNA-protein interactions; Nanobody; CUT&Tag; Epigenomics; Tn5 tagmentation; Chromatin profiling
Organisational unit
03518 - Mansuy, Isabelle / Mansuy, Isabelle
Notes
Funding
ETH-10 15-2 - Long non-coding-RNA's: Study theirpotential role as novel regulators of brain functions across generations" (ETHZ)
182880 - NCCR RNA#38;Disease (51NF40-182880): Flexibility Grant (SNF)
20-1 FEL-28 - Study of the 3D neuronal genome in experience-dependent brain functions. (ETHZ)
182880 - NCCR RNA#38;Disease (51NF40-182880): Flexibility Grant (SNF)
20-1 FEL-28 - Study of the 3D neuronal genome in experience-dependent brain functions. (ETHZ)