Merlin: Computer-Aided Oligonucleotide Design for Large Scale Genome Engineering with MAGE.

Abstract:

:Genome engineering technologies now enable precise manipulation of organism genotype, but can be limited in scalability by their design requirements. Here we describe Merlin ( http://merlincad.org ), an open-source web-based tool to assist biologists in designing experiments using multiplex automated genome engineering (MAGE). Merlin provides methods to generate pools of single-stranded DNA oligonucleotides (oligos) for MAGE experiments by performing free energy calculation and BLAST scoring on a sliding window spanning the targeted site. These oligos are designed not only to improve recombination efficiency, but also to minimize off-target interactions. The application further assists experiment planning by reporting predicted allelic replacement rates after multiple MAGE cycles, and enables rapid result validation by generating primer sequences for multiplexed allele-specific colony PCR. Here we describe the Merlin oligo and primer design procedures and validate their functionality compared to OptMAGE by eliminating seven AvrII restriction sites from the Escherichia coli genome.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Quintin M,Ma NJ,Ahmed S,Bhatia S,Lewis A,Isaacs FJ,Densmore D

doi

10.1021/acssynbio.5b00219

subject

Has Abstract

pub_date

2016-06-17 00:00:00

pages

452-8

issue

6

issn

2161-5063

journal_volume

5

pub_type

杂志文章
  • Engineering Prokaryotic Transcriptional Activator XylR as a Xylose-Inducible Biosensor for Transcription Activation in Yeast.

    abstract::Biosensors regulated by specific substrates are needed to develop genetic tools to meet the needs of engineering microbial cell factories. Here, a xylose-inducible biosensor (xylbiosensor), comprising the Escherichia coli activation factor XylR, fusion activation domain (AD) VPRH, and a hybrid promoter with operator x...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00122

    authors: Wei W,Shang Y,Zhang P,Liu Y,You D,Yin B,Ye B

    更新日期:2020-05-15 00:00:00

  • Efficient and Precise Genome Editing in Shewanella with Recombineering and CRISPR/Cas9-Mediated Counter-Selection.

    abstract::Dissimilatory metal-reducing bacteria, particularly those from the genus Shewanella, are of importance for bioremediation of metal contaminated sites and sustainable energy production. However, studies on this species have suffered from a lack of effective genetic tools for precise and high throughput genome manipulat...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00188

    authors: Corts AD,Thomason LC,Gill RT,Gralnick JA

    更新日期:2019-08-16 00:00:00

  • Rationalizing Context-Dependent Performance of Dynamic RNA Regulatory Devices.

    abstract::The ability of RNA to sense, regulate, and store information is an attractive attribute for a variety of functional applications including the development of regulatory control devices for synthetic biology. RNA folding and function is known to be highly context sensitive, which limits the modularity and reuse of RNA ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00041

    authors: Kent R,Halliwell S,Young K,Swainston N,Dixon N

    更新日期:2018-07-20 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

  • 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

  • Exploiting Spermidine N-Hydroxycinnamoyltransferase Diversity and Substrate Promiscuity to Produce Various Trihydroxycinnamoyl Spermidines and Analogues in Engineered Yeast.

    abstract::Trihydroxycinnamoyl spermidines (THCSpd) are plant specialized metabolites with promising pharmacological activities as antifungals, antibacterial, antiviral, and antidepressant drugs. However, their characterization and potential pharmaceutical exploitation are greatly impaired by the sourcing of these compounds, res...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00391

    authors: Perrin J,Kulagina N,Unlubayir M,Munsch T,Carqueijeiro I,Dugé de Bernonville T,De Craene JO,Clastre M,St-Pierre B,Giglioli-Guivarc'h N,Gagneul D,Lanoue A,Courdavault V,Besseau S

    更新日期:2021-01-15 00:00:00

  • Systematic Tools for Reprogramming Plant Gene Expression in a Simple Model, Marchantia polymorpha.

    abstract::We present the OpenPlant toolkit, a set of interlinked resources and techniques to develop Marchantia as testbed for bioengineering in plants. Marchantia is a liverwort, a simple plant with an open form of development that allows direct visualization of gene expression and dynamics of cellular growth in living tissues...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00511

    authors: Sauret-Güeto S,Frangedakis E,Silvestri L,Rebmann M,Tomaselli M,Markel K,Delmans M,West A,Patron NJ,Haseloff J

    更新日期:2020-04-17 00:00:00

  • Mathematical Modeling of RNA-Based Architectures for Closed Loop Control of Gene Expression.

    abstract::Feedback allows biological systems to control gene expression precisely and reliably, even in the presence of uncertainty, by sensing and processing environmental changes. Taking inspiration from natural architectures, synthetic biologists have engineered feedback loops to tune the dynamics and improve the robustness ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00040

    authors: Agrawal DK,Tang X,Westbrook A,Marshall R,Maxwell CS,Lucks J,Noireaux V,Beisel CL,Dunlop MJ,Franco E

    更新日期:2018-05-18 00:00:00

  • Synthetic Derivatives of (+)-epi-α-Bisabolol Are Formed by Mammalian Cytochromes P450 Expressed in a Yeast Reconstituted Pathway.

    abstract::Identification of the enzyme(s) involved in complex biosynthetic pathways can be challenging. An alternative approach might be to deliberately diverge from the original natural enzyme source and use promiscuous enzymes from other organisms. In this paper, we have tested the ability of a series of human and animal cyto...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00399

    authors: Sarrade-Loucheur A,Ro DK,Fauré R,Remaud-Siméon M,Truan G

    更新日期:2020-02-21 00:00:00

  • Characterization and modeling of transcriptional cross-regulation in Acinetobacter baylyi ADP1.

    abstract::Synthetic biology involves reprogramming and engineering of regulatory genes in innovative ways for the implementation of novel tasks. Transcriptional gene regulation systems induced by small molecules in prokaryotes provide a rich source for logic gates. Cross-regulation, whereby a promoter is activated by different ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb3000244

    authors: Zhang D,Zhao Y,He Y,Wang Y,Zhao Y,Zheng Y,Wei X,Zhang L,Li Y,Jin T,Wu L,Wang H,Davison PA,Xu J,Huang WE

    更新日期:2012-07-20 00:00:00

  • Lessons from Two Design-Build-Test-Learn Cycles of Dodecanol Production in Escherichia coli Aided by Machine Learning.

    abstract::The Design-Build-Test-Learn (DBTL) cycle, facilitated by exponentially improving capabilities in synthetic biology, is an increasingly adopted metabolic engineering framework that represents a more systematic and efficient approach to strain development than historical efforts in biofuels and biobased products. Here, ...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00020

    authors: Opgenorth P,Costello Z,Okada T,Goyal G,Chen Y,Gin J,Benites V,de Raad M,Northen TR,Deng K,Deutsch S,Baidoo EEK,Petzold CJ,Hillson NJ,Garcia Martin H,Beller HR

    更新日期:2019-06-21 00:00:00

  • Bringing Light into Cell-Free Expression.

    abstract::Cell-free systems, as part of the synthetic biology field, have become a critical platform in biological studies. However, there is a lack of research into developing a switch for a dynamical control of the transcriptional and translational process. The optogenetic tool has been widely proven as an ideal control switc...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00211

    authors: Zhang P,Yang J,Cho E,Lu Y

    更新日期:2020-08-21 00:00:00

  • Development of a Transcription Factor-Based Lactam Biosensor.

    abstract::Lactams are an important class of commodity chemicals used in the manufacture of nylons, with millions of tons produced every year. Biological production of lactams could be greatly improved by high-throughput sensors for lactam biosynthesis. To identify biosensors of lactams, we applied a chemoinformatic approach ins...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00136

    authors: Zhang J,Barajas JF,Burdu M,Ruegg TL,Dias B,Keasling JD

    更新日期:2017-03-17 00:00:00

  • Strategies for Editing Virulent Staphylococcal Phages Using CRISPR-Cas10.

    abstract::Staphylococci are prevalent skin-dwelling bacteria that are also leading causes of antibiotic-resistant infections. Viruses that infect and lyse these organisms (virulent staphylococcal phages) can be used as alternatives to conventional antibiotics and represent promising tools to eliminate or manipulate specific spe...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00240

    authors: Bari SMN,Walker FC,Cater K,Aslan B,Hatoum-Aslan A

    更新日期:2017-12-15 00:00:00

  • Metabolic Engineering of Escherichia coli for Efficient Production of 2-Pyrone-4,6-dicarboxylic Acid from Glucose.

    abstract::2-Pyrone-4,6-dicarboxylic acid (PDC) is a pseudoaromatic dicarboxylic acid and is a promising biobased building block chemical that can be used to make diverse polyesters with novel functionalities. In this study, Escherichia coli was metabolically engineered to produce PDC from glucose. First, an efficient biosynthet...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00281

    authors: Luo ZW,Kim WJ,Lee SY

    更新日期:2018-09-21 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

  • Conditional Recruitment to a DNA-Bound CRISPR-Cas Complex Using a Colocalization-Dependent Protein Switch.

    abstract::To spatially control biochemical functions at specific sites within a genome, we have engineered a synthetic switch that activates when bound to its DNA target site. The system uses two CRISPR-Cas complexes to colocalize components of a de novo-designed protein switch (Co-LOCKR) to adjacent sites in the genome. Coloca...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.0c00012

    authors: Kirkpatrick RL,Lewis K,Langan RA,Lajoie MJ,Boyken SE,Eakman M,Baker D,Zalatan JG

    更新日期:2020-09-18 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

  • The Resistome: A Comprehensive Database of Escherichia coli Resistance Phenotypes.

    abstract::The microbial ability to resist stressful environmental conditions and chemical inhibitors is of great industrial and medical interest. Much of the data related to mutation-based stress resistance, however, is scattered through the academic literature, making it difficult to apply systematic analyses to this wealth of...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00150

    authors: Winkler JD,Halweg-Edwards AL,Erickson KE,Choudhury A,Pines G,Gill RT

    更新日期:2016-12-16 00:00:00

  • Sequence Specific Modeling of E. coli Cell-Free Protein Synthesis.

    abstract::Cell-free protein synthesis (CFPS) is a widely used research tool in systems and synthetic biology. However, if CFPS is to become a mainstream technology for applications such as point of care manufacturing, we must understand the performance limits and costs of these systems. Toward this question, we used sequence sp...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.7b00465

    authors: Vilkhovoy M,Horvath N,Shih CH,Wayman JA,Calhoun K,Swartz J,Varner JD

    更新日期:2018-08-17 00:00:00

  • Engineering Synthetic Proteins to Generate Ca2+ Signals in Mammalian Cells.

    abstract::The versatility of Ca2+ signals allows it to regulate diverse cellular processes such as migration, apoptosis, motility and exocytosis. In some receptors (e.g., VEGFR2), Ca2+ signals are generated upon binding their ligand(s) (e.g., VEGF-A). Here, we employed a design strategy to engineer proteins that generate a Ca2+...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.6b00310

    authors: Qudrat A,Truong K

    更新日期:2017-03-17 00:00:00

  • Active Analyte Import Improves the Dynamic Range and Sensitivity of a Vitamin B12 Biosensor.

    abstract::Cell-free systems provide a versatile platform for the development of low-cost, easy-to-use sensors for diverse analytes. However, sensor affinity dictates response sensitivity, and improving binding affinity can be challenging. Here, we describe efforts to address this problem while developing a biosensor for vitamin...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.9b00429

    authors: McNerney MP,Piorino F,Michel CL,Styczynski MP

    更新日期:2020-02-21 00:00:00

  • Boolean Computation in Plants Using Post-translational Genetic Control and a Visual Output Signal.

    abstract::Due to autotrophic growing capacity and extremely rich secondary metabolism, plants should be preferred targets of synthetic biology. However, developments in plants usually run below those in other taxonomic groups. In this work we engineered genetic circuits capable of logic YES, OR and AND Boolean computation in pl...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00214

    authors: Cordero T,Rosado A,Majer E,Jaramillo A,Rodrigo G,Daròs JA

    更新日期:2018-10-19 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

  • Semisupervised Gaussian Process for Automated Enzyme Search.

    abstract::Synthetic biology is today harnessing the design of novel and greener biosynthesis routes for the production of added-value chemicals and natural products. The design of novel pathways often requires a detailed selection of enzyme sequences to import into the chassis at each of the reaction steps. To address such desi...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.5b00294

    authors: Mellor J,Grigoras I,Carbonell P,Faulon JL

    更新日期:2016-06-17 00:00:00

  • Tn-Core: A Toolbox for Integrating Tn-seq Gene Essentiality Data and Constraint-Based Metabolic Modeling.

    abstract::The design of synthetic cells requires a detailed understanding of the relevance of genes and gene networks underlying complex cellular phenotypes. Transposon-sequencing (Tn-seq) and constraint-based metabolic modeling can be used to probe the core genetic and metabolic networks underlying a biological process. Integr...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/acssynbio.8b00432

    authors: diCenzo GC,Mengoni A,Fondi M

    更新日期:2019-01-18 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

  • Programming controlled adhesion of E. coli to target surfaces, cells, and tumors with synthetic adhesins.

    abstract::In this work we report synthetic adhesins (SAs) enabling the rational design of the adhesion properties of E. coli. SAs have a modular structure comprising a stable β-domain for outer membrane anchoring and surface-exposed immunoglobulin domains with high affinity and specificity that can be selected from large repert...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb500252a

    authors: Piñero-Lambea C,Bodelón G,Fernández-Periáñez R,Cuesta AM,Álvarez-Vallina L,Fernández LÁ

    更新日期:2015-04-17 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

  • Evolutionary stability of a refactored phage genome.

    abstract::Engineered genetic systems are commonly unstable; if propagated, they evolve to reverse or modify engineered elements because the elements impair fitness. A goal of synthetic biology is thus to anticipate and avoid detrimental engineering, but little is yet known about which types of elements cause problems in differe...

    journal_title:ACS synthetic biology

    pub_type: 杂志文章

    doi:10.1021/sb300040v

    authors: Springman R,Molineux IJ,Duong C,Bull RJ,Bull JJ

    更新日期:2012-09-21 00:00:00