A Scalable Epitope Tagging Approach for High Throughput ChIP-Seq Analysis.

Abstract:

:Eukaryotic transcriptional factors (TFs) typically recognize short genomic sequences alone or together with other proteins to modulate gene expression. Mapping of TF-DNA interactions in the genome is crucial for understanding the gene regulatory programs in cells. While chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is commonly used for this purpose, its application is severely limited by the availability of suitable antibodies for TFs. To overcome this limitation, we developed an efficient and scalable strategy named cmChIP-Seq that combines the clustered regularly interspaced short palindromic repeats (CRISPR) technology with microhomology mediated end joining (MMEJ) to genetically engineer a TF with an epitope tag. We demonstrated the utility of this tool by applying it to four TFs in a human colorectal cancer cell line. The highly scalable procedure makes this strategy ideal for ChIP-Seq analysis of TFs in diverse species and cell types.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Xiong X,Zhang Y,Yan J,Jain S,Chee S,Ren B,Zhao H

doi

10.1021/acssynbio.6b00358

subject

Has Abstract

pub_date

2017-06-16 00:00:00

pages

1034-1042

issue

6

issn

2161-5063

journal_volume

6

pub_type

杂志文章
  • Reversing an Extracellular Electron Transfer Pathway for Electrode-Driven Acetoin Reduction.

    abstract::Microbial electrosynthesis is an emerging technology with the potential to simultaneously store renewably generated energy, fix carbon dioxide, and produce high-value organic compounds. However, limited understanding of the route of electrons into the cell remains an obstacle to developing a robust microbial electrosy...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00498

    authors: Tefft NM,TerAvest MA

    更新日期:2019-07-19 00:00:00

  • Systematic Identification of a Panel of Strong Constitutive Promoters from Streptomyces albus.

    abstract::Actinomycetes are important organisms for the biosynthesis of valuable natural products. However, only a limited number of well-characterized native constitutive promoters from actinomycetes are available for the construction and engineering of large biochemical pathways. Here, we report the discovery and characteriza...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00016

    authors: Luo Y,Zhang L,Barton KW,Zhao H

    更新日期:2015-09-18 00:00:00

  • Synthetic Biogenesis of Bacterial Amyloid Nanomaterials with Tunable Inorganic-Organic Interfaces and Electrical Conductivity.

    abstract::Amyloids are highly ordered, hierarchal protein nanoassemblies. Functional amyloids in bacterial biofilms, such as Escherichia coli curli fibers, are formed by the polymerization of monomeric proteins secreted into the extracellular space. Curli is synthesized by living cells, is primarily composed of the major curlin...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00166

    authors: Seker UO,Chen AY,Citorik RJ,Lu TK

    更新日期:2017-02-17 00:00:00

  • Switching Protein Localization by Site-Directed RNA Editing under Control of Light.

    abstract::Site directed RNA editing is an engineered tool for the posttranscriptional manipulation of RNA and proteins. Here, we demonstrate the inclusion of additional N- and C-terminal protein domains in an RNA editing-dependent manner to switch between protein isoforms in mammalian cell culture. By inclusion of localization ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00113

    authors: Vogel P,Hanswillemenke A,Stafforst T

    更新日期:2017-09-15 00:00:00

  • Assembly of Multicomponent Protein Filaments Using Engineered Subunit Interfaces.

    abstract::Exploiting the ability of proteins to self-assemble into architectural templates may provide novel routes for the positioning of functional molecules in nanotechnology. Here we report the engineering of multicomponent protein templates composed of distinct monomers that assemble in repeating orders into a dynamic func...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00241

    authors: Glover DJ,Lim S,Xu D,Sloan NB,Zhang Y,Clark DS

    更新日期:2018-10-19 00:00:00

  • An Artificial Yeast Genetic Circuit Enables Deep Mutational Scanning of an Antimicrobial Resistance Protein.

    abstract::Understanding the molecular mechanisms underlying antibiotic resistance requires concerted efforts in enzymology and medicinal chemistry. Here we describe a new synthetic biology approach to antibiotic development, where the presence of tetracycline antibiotics is linked to a life-death selection in Saccharomyces cere...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00121

    authors: Scott LH,Mathews JC,Flematti GR,Filipovska A,Rackham O

    更新日期:2018-08-17 00:00:00

  • Design and Selection of a Synthetic Feedback Loop for Optimizing Biofuel Tolerance.

    abstract::Feedback control allows cells to dynamically sense and respond to environmental changes. However, synthetic controller designs can be challenging because of implementation issues, such as determining optimal expression levels for circuit components within a feedback loop. Here, we addressed this by coupling rational d...

    journal_title:ACS synthetic biology

    pub_type: 信件

    doi:10.1021/acssynbio.7b00260

    authors: Siu Y,Fenno J,Lindle JM,Dunlop MJ

    更新日期:2018-01-19 00:00:00

  • Genome Engineering of Virulent Lactococcal Phages Using CRISPR-Cas9.

    abstract::Phages are biological entities found in every ecosystem. Although much has been learned about them in past decades, significant knowledge gaps remain. Manipulating virulent phage genomes is challenging. To date, no efficient gene-editing tools exist for engineering virulent lactococcal phages. Lactococcus lactis is a ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00388

    authors: Lemay ML,Tremblay DM,Moineau S

    更新日期:2017-07-21 00:00:00

  • 3D Printing for the Fabrication of Biofilm-Based Functional Living Materials.

    abstract::Bacterial biofilms are three-dimensional networks of cells entangled in a self-generated extracellular polymeric matrix composed of proteins, lipids, polysaccharides, and nucleic acids. Biofilms can establish themselves on virtually any accessible surface and lead to varying impacts ranging from infectious diseases to...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00192

    authors: Balasubramanian S,Aubin-Tam ME,Meyer AS

    更新日期:2019-07-19 00:00:00

  • Microbial Synthesis of Human-Hormone Melatonin at Gram Scales.

    abstract::Melatonin is a commercially attractive tryptophan-derived hormone. Here we describe a bioprocess for the production of melatonin using Escherichia coli to high titers. The first engineered strain produced 0.13 g/L of melatonin from tryptophan under fed-batch fermentation conditions. A 4-fold improvement on melatonin t...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00065

    authors: Luo H,Schneider K,Christensen U,Lei Y,Herrgard M,Palsson BØ

    更新日期:2020-06-19 00:00:00

  • An Engineered Survival-Selection Assay for Extracellular Protein Expression Uncovers Hypersecretory Phenotypes in Escherichia coli.

    abstract::The extracellular expression of recombinant proteins using laboratory strains of Escherichia coli is now routinely achieved using naturally secreted substrates, such as YebF or the osmotically inducible protein Y (OsmY), as carrier molecules. However, secretion efficiency through these pathways needs to be improved fo...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00366

    authors: Natarajan A,Haitjema CH,Lee R,Boock JT,DeLisa MP

    更新日期:2017-05-19 00:00:00

  • Artificial conversion of the mating-type of Saccharomyces cerevisiae without autopolyploidization.

    abstract::Crossbreeding is a classical yeast hybridization procedure, where the mating of haploid cells of opposite mating-type, MATa and MATα cells, produces a new heterozygous diploid. Here, we describe a method to generate haploid MATa and MATα cells using mating-type conversion caused by expression of the HO gene, which enc...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400016j

    authors: Fukuda N,Matsukura S,Honda S

    更新日期:2013-12-20 00:00:00

  • Single Day Construction of Multigene Circuits with 3G Assembly.

    abstract::The ability to rapidly design, build, and test prototypes is of key importance to every engineering discipline. DNA assembly often serves as a rate limiting step of the prototyping cycle for synthetic biology. Recently developed DNA assembly methods such as isothermal assembly and type IIS restriction enzyme systems t...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00060

    authors: Halleran AD,Swaminathan A,Murray RM

    更新日期:2018-05-18 00:00:00

  • Engineered Photoactivatable Genetic Switches Based on the Bacterium Phage T7 RNA Polymerase.

    abstract::Genetic switches in which the activity of T7 RNA polymerase (RNAP) is directly regulated by external signals are obtained with an engineering strategy of splitting the protein into fragments and using regulatory domains to modulate their reconstitutions. Robust switchable systems with excellent dark-off/light-on prope...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00248

    authors: Han T,Chen Q,Liu H

    更新日期:2017-02-17 00:00:00

  • The Formal Language and Design Principles of Autonomous DNA Walker Circuits.

    abstract::Simple computation can be performed using the interactions between single-stranded molecules of DNA. These interactions are typically toehold-mediated strand displacement reactions in a well-mixed solution. We demonstrate that a DNA circuit with tethered reactants is a distributed system and show how it can be describ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00275

    authors: Boemo MA,Lucas AE,Turberfield AJ,Cardelli L

    更新日期:2016-08-19 00:00:00

  • Overcoming the Challenges of Megabase-Sized Plasmid Construction in Escherichia coli.

    abstract::Although Escherichia coli has been a popular tool for plasmid construction, this bacterium was believed to be "unsuitable" for constructing a large plasmid whose size exceeds 500 kilobases. We assumed that traditional plasmid vectors may lack some regulatory DNA elements required for the stable replication and segrega...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00008

    authors: Mukai T,Yoneji T,Yamada K,Fujita H,Nara S,Su'etsugu M

    更新日期:2020-06-19 00:00:00

  • Enhancing Intercellular Coordination: Rewiring Quorum Sensing Networks for Increased Protein Expression through Autonomous Induction.

    abstract::While inducing agents are often used to redirect resources from growth and proliferation toward product outputs, they can be prohibitively expensive on the industrial scale. Previously, we developed an autonomously guided protein production system based on the rewiring of E. coli's native quorum sensing (QS) signal tr...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00261

    authors: Zargar A,Quan DN,Bentley WE

    更新日期:2016-09-16 00:00:00

  • Cooperativity to increase Turing pattern space for synthetic biology.

    abstract::It is hard to bridge the gap between mathematical formulations and biological implementations of Turing patterns, yet this is necessary for both understanding and engineering these networks with synthetic biology approaches. Here, we model a reaction-diffusion system with two morphogens in a monostable regime, inspire...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb500233u

    authors: Diambra L,Senthivel VR,Menendez DB,Isalan M

    更新日期:2015-02-20 00:00:00

  • Circumvention of Learning Increases Intoxication Efficacy of Nematicidal Engineered Bacteria.

    abstract::Synthetic biology holds promise to engineer systems to treat diseases. One critical, yet underexplored, facet of designing such systems is the interplay between the system and the pathogen. Understanding this interplay may be critical to increasing efficacy and overcoming resistance against the system. Using the princ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00192

    authors: Bracho OR,Manchery C,Haskell EC,Blanar CA,Smith RP

    更新日期:2016-03-18 00:00:00

  • Engineering Escherichia coli into a protein delivery system for mammalian cells.

    abstract::Many Gram-negative pathogens encode type 3 secretion systems, sophisticated nanomachines that deliver proteins directly into the cytoplasm of mammalian cells. These systems present attractive opportunities for therapeutic protein delivery applications; however, their utility has been limited by their inherent pathogen...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00002

    authors: Reeves AZ,Spears WE,Du J,Tan KY,Wagers AJ,Lesser CF

    更新日期:2015-05-15 00:00:00

  • Rational Design of the N-Terminal Coding Sequence for Regulating Enzyme Expression in Bacillus subtilis.

    abstract::Synonymous mutation of the N-terminal coding sequence (NCS) has been used to regulate gene expression. We here developed a statistical model to predict the effect of the NCSs on protein expression in Bacillus subtilis WB600. First, a synonymous mutation was performed within the first 10 residues of a superfolder green...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00309

    authors: Xu K,Tong Y,Li Y,Tao J,Li J,Zhou J,Liu S

    更新日期:2021-01-19 00:00:00

  • Enhanced triterpene accumulation in Panax ginseng hairy roots overexpressing mevalonate-5-pyrophosphate decarboxylase and farnesyl pyrophosphate synthase.

    abstract::To elucidate the function of mevalonate-5-pyrophosphate decarboxylase (MVD) and farnesyl pyrophosphate synthase (FPS) in triterpene biosynthesis, the genes governing the expression of these enzymes were transformed into Panax ginseng hairy roots. All the transgenic lines showed higher expression levels of PgMVD and Pg...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400194g

    authors: Kim YK,Kim YB,Uddin MR,Lee S,Kim SU,Park SU

    更新日期:2014-10-17 00:00:00

  • Biophysical, mutational, and functional investigation of the chromophore-binding pocket of light-oxygen-voltage photoreceptors.

    abstract::As light-regulated actuators, sensory photoreceptors underpin optogenetics and numerous applications in synthetic biology. Protein engineering has been applied to fine-tune the properties of photoreceptors and to generate novel actuators. For the blue-light-sensitive light-oxygen-voltage (LOV) photoreceptors, mutation...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400205x

    authors: Diensthuber RP,Engelhard C,Lemke N,Gleichmann T,Ohlendorf R,Bittl R,Möglich A

    更新日期:2014-11-21 00:00:00

  • The spinach RNA aptamer as a characterization tool for synthetic biology.

    abstract::Characterization of genetic control elements is essential for the predictable engineering of synthetic biology systems. The current standard for in vivo characterization of control elements is through the use of fluorescent reporter proteins such as green fluorescent protein (GFP). Gene expression, however, involves n...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400089c

    authors: Pothoulakis G,Ceroni F,Reeve B,Ellis T

    更新日期:2014-03-21 00:00:00

  • Effects of compartment size on the kinetics of intracompartmental multimeric protein synthesis.

    abstract::The cell contents are encapsulated within a compartment, the volume of which is a fundamental physical parameter that may affect intracompartmental reactions. However, there have been few studies to elucidate whether and how volume changes alone can affect the reaction kinetics. It is difficult to address these questi...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb300041z

    authors: Matsuura T,Hosoda K,Kazuta Y,Ichihashi N,Suzuki H,Yomo T

    更新日期:2012-09-21 00:00:00

  • Electrochemical Measurement of the β-Galactosidase Reporter from Live Cells: A Comparison to the Miller Assay.

    abstract::In order to match our ability to conceive of and construct cells with enhanced function, we must concomitantly develop facile, real-time methods for elucidating performance. With these, new designs can be tested in silico and steps in construction incrementally validated. Electrochemical monitoring offers the above ad...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00073

    authors: Tschirhart T,Zhou XY,Ueda H,Tsao CY,Kim E,Payne GF,Bentley WE

    更新日期:2016-01-15 00:00:00

  • Multiplex iterative plasmid engineering for combinatorial optimization of metabolic pathways and diversification of protein coding sequences.

    abstract::Engineering complex biological systems typically requires combinatorial optimization to achieve the desired functionality. Here, we present Multiplex Iterative Plasmid Engineering (MIPE), which is a highly efficient and customized method for combinatorial diversification of plasmid sequences. MIPE exploits ssDNA media...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb400051t

    authors: Li Y,Gu Q,Lin Z,Wang Z,Chen T,Zhao X

    更新日期:2013-11-15 00:00:00

  • Transcription activator-like effectors: a toolkit for synthetic biology.

    abstract::Transcription activator-like effectors (TALEs) are proteins secreted by Xanthomonas bacteria to aid the infection of plant species. TALEs assist infections by binding to specific DNA sequences and activating the expression of host genes. Recent results show that TALE proteins consist of a central repeat domain, which ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章,评审

    doi:10.1021/sb400137b

    authors: Moore R,Chandrahas A,Bleris L

    更新日期:2014-10-17 00:00:00

  • Exploring Chemical Biosynthetic Design Space with Transform-MinER.

    abstract::Transform-MinER (Transforming Molecules in Enzyme Reactions) is a web application facilitating the exploration of chemical biosynthetic space, guiding the user toward promising start points for enzyme design projects or directed evolution experiments. Two types of search are possible: Molecule Search allows a user to ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00105

    authors: Tyzack JD,Ribeiro AJM,Borkakoti N,Thornton JM

    更新日期:2019-11-15 00:00:00

  • Specification and simulation of synthetic multicelled behaviors.

    abstract::Recent advances in the design and construction of synthetic multicelled systems in E. coli and S. cerevisiae suggest that it may be possible to implement sophisticated distributed algorithms with these relatively simple organisms. However, existing design frameworks for synthetic biology do not account for the unique ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb300034m

    authors: Jang SS,Oishi KT,Egbert RG,Klavins E

    更新日期:2012-08-17 00:00:00