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... two global stock centers for Arabidopsis thaliana (Arabidopsis). Our mission is to collect, preserve,...in 60 countries each year. Introducing Arabidopsis Arabidopsis is an important model system for research...and you have yourself a plant publicity crisis. The Arabidopsis Biological Resource Center (ABRC) set out...Translating Research on Arabidopsis Into a Network of Educational Resources). Arabidopsis is used in research... the Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for...Ohio State University. Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two...distribute seeds, DNA and other resources for Arabidopsis and related species. Located at The Ohio State...

...spectinomycin resistant phenotype in bacteria. While there are many known spectinomycin resistance mutations...genomic site for mutagenesis, a feature which they used to generate bacteria resistant to both spectinomycin...directed mutagenesis to generate specific mutations in your plasmids Use REPLACER Mutagenesis to mutate...Mutagenesis is a tool that both evolution and molecular biologists use to tinker with DNA. Making changes...region. EvolvR, a CRISPR-Cas9 based targeted mutagenesis method developed by the Dueber Lab at Berkeley...potential applications. Challenges of existing mutagenesis methods Several continuous diversification methods...error prone polymerase I (PolI3M) is capable of mutagenesis in E. Coli, but this approach requires special...

..., a graduate student at Stanford University. Arabidopsis is a fantastic model organism for many reasons... are many motivations to generate transgenic Arabidopsis, from studying transcriptional and translational... plants, to complementing mutant phenotypes. Arabidopsis is amenable to the floral drip or dip transformation...transformation protocol, as well as protocols for growing Arabidopsis are also available (Weigel and Glazebrook, 2002...vector is the first step in generating transgenic Arabidopsis. There are a bunch of great options, but here... pool you will be selecting from later. The Arabidopsis transformation process The process itself is ... the chances of tDNA integration. Selecting Arabidopsis transformants Once your seeds from your T0 plants...

... the mutagenesis reaction. Alternative approaches to site directed mutagenesis Point-mutagenesis is fairly...constructs), in functional analysis of proteins (to perform alanine scanning mutagenesis or targeted substitution...should be resistant to Cas9 cleavage. If a plasmid contains the template, site-directed mutagenesis can be...resulting construct resistant to Cas9 induced cleavage. Summary of Site Directed Mutagenesis In brief, point-mutations...mutation of interest. Gel electrophoresis diagrams of the anticipated restriction analysis results (left) and...Laursen from Cornell University. Site directed mutagenesis is a highly versatile technique that can be used...substitution of key residues), and in SNP analysis (to introduce naturally occuring SNPs in a plasmid context...

...protein synthesis Bactericidal 25-100 ug/mL Ampicillin beta-lactam Inhibits cell wall synthesis Bactericidal...Resistance to antibiotics is a widely used tool in molecular biology, yet scientists rarely stop to think...to maintain the plasmid. Adding an antibiotic resistance gene to the plasmid solves both problems at once...pressure to keep your plasmid. Viva la (bacterial) resistance!  What are antibiotics? Antibiotics are generally...Binds 30S ribosomal subunit; interrupts protein synthesis Bactericidal 7.5-50 ug/mL Streptomycin aminoglycoside...Carbenicillin beta-lactam Inhibits cell wall synthesis Bactericidal 100 ug/mL Bleomycin glycopeptide...Binds 30S ribosomal subunit; inhibits protein synthesis (elongation step) Bacteriostatic 10 ug/mL ...

...Microbiology Pages What is antibiotic resistance? Antibiotic resistance is defined as microbial survival despite...generally organisms with multi-drug resistance are resistant to three or more different classes, or types...Learn How Antibiotic Resistance Genes Are Used in the Lab How does antibiotic resistance develop? There are...in which bacteria can become resistant to antibacterial drugs. Resistance can be intrinsic, meaning a ...means the bacterium is not naturally resistant but has become resistant somehow overtime. One fantastic example... example of intrinsic resistance would be Mycoplasma species’ resistance to beta-lactam antibiotics, like...Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired resistance can be the result of ...

...this insertion size, primer synthesis is more expensive than gBlock synthesis, and multifragment assembly...REPLACR-mutagenesis could save busy scientists a lot of time. Let us know how REPLACR-mutagenesis works ...Comparison of REPLACR-mutagenesis with other common methods for site directed mutagenesis   References 1. ...al. (2016). “REPLACR-mutagenesis: a one-step method for site-directed mutagenesis by recombineering.” ...Site-directed mutagenesis (SDM) is one of the key tools researchers use to prove causation in molecular...vitro assembly methods. A new method, REPLACR-mutagenesis, harnesses the power of bacterial recombineering... Plasmids After PCR). Current site directed mutagenesis methods Current SDM methods rely on in vitro ...

...beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces beta-lactamase to ...processes (e.g., protein synthesis), while amp only affects the cell wall synthesis step during cell division...aminoglycoside which inhibits protein synthesis. The kan resistance gene, NPTII, facilitates phosphorylation...cells, are already resistant to this antibiotic due to stable integration of resistance genes in their genomes...damage. Choosing an antibiotic resistance Choosing the best antibiotic resistance ultimately comes down to ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...interest. But does it matter which antibiotic resistance gene you select? In this blog, we’ll break down...

...interest, C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants....Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle work in any plant species...Versatile and Efficient Cloning System for Plant Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013)...

...Apoptosis or “programmed cell death" plays a pivotal role in an array of biological processes including...development, the immune system, and cell turnover. Apoptosis is a highly controlled process that is triggered...fragmentation and death (Elmore, 2007). To study apoptosis in vivo, scientists have used genetically encoded...fluorescent executioner caspase reporters. Several apoptosis reporters have been developed, such as the Forster...Zhang et al., 2019). Thus to effectively study apoptosis in vivo scientists need a protease reporter that...brightness. The Shu Lab from UCSF designed an improved apoptosis reporter, ZipGFP, which utilized the self-assembling... better study the spatio-temporal dynamics of apoptosis in living animals. To do this they modified a ...

...effects. One-pot saturation mutagenesis The one-pot saturation mutagenesis technique described in Wrenbeck... as the gene or use custom gene synthesis or site directed mutagenesis to remove the BbvCI site(s). ...saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How .... “Random Mutagenesis by Error-Prone PCR”. Methods in Molecular Biology In Vitro Mutagenesis Protocols...the Whitehead lab developed One pot saturation mutagenesis as a quick and easy technique that can be used...library generation prior to one pot saturation mutagenesis Several methods can be used to create mutational... shortcomings compared to one pot saturation mutagenesis. Error-prone PCR: This method amplifies the...

...take a look at our live examples: experiment or analysis. In the rest of this article, we'll explain the...batch guide design, batch primer design or batch analysis. We wanted to relieve scientists of the burden... by Luca Pinello and Kendell Clement for batch analysis because of its ease-of-use and comprehensive reports...publicly-available for high-throughput design and analysis of CRISPR experiments. It has certainly boosted...

...research on diseases of glycosylation, arginine mutagenesis, and neural development. Paula Videira: Congenital...disorders of glycosylation Paula Videira is an Assistant Professor at the Faculty of Sciences of NOVA University...functional activity of VUSs will enable more accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith...

...plasmids. Here, we'll cover restriction digest analysis. Diagnostic restriction digest Diagnostic digests...different features within the plasmid. Restriction analysis can also be used successfully even if you don'...

... more long-term solution is the use of unique persistent identifiers (PIDs), a long-lasting way to identify...documents, files, or physical reagents. What are persistent identifiers? Perhaps the most commonly known ...original article online. It is also designed to be persistent and exist online indefinitely. PIDs historically...plasmids! In an effort to support standardized persistent identification of research reagents, several ...this post and sharing her technical expertise in persistent identifiers. Additional Resources on the Addgene...

...Mycobacterium tuberculosis resistance to pyrazinamide (PZA) (a drug commonly used to treat tuberculosis) and in...projects.   Understanding drug resistance mechanisms of Mycobacterium tuberculosis Dr. Patricia Sheen, Universidad... accurate detection of PZA-resistance is necessary to improve tuberculosis care and control. In her current...methods and improved therapies for PZA-resistant Mycobacteria tuberculosis. Investigation of multispecies biofilm...Sheen aims to identify new genes related to PZA-resistance by using different approaches, including CRISPR...experience in the field of bacterial and fungal pathogenesis. Her research focuses on a collection of pathogenic...

... Protocols Sequence Analysis Sequence Analysis of a Plasmid You ...performing sequence analysis of a plasmid Protocols...

...Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Cell. 2014...transcriptional activation of endogenous genes and it consists of three components: 1. A nucleolytically inactive... a synthetic transcription activation complex consisting of multiple distinct effector domains that work...genes that, when turned on, make melanoma cells resistant to treatment with a BRAF inhibitor. (The test ...-mediated screens exhibited a high degree of consistency and validation with >80% effectively enriched...

...only measures infectious viral particles. One of the si mplest methods of measuring functional titer uses...components such as LTRs, gag, WPRE, antibiotic resistance-genes, or the transgene itself. Many researchers...Another popular approach utilizes the antibiotic resistance genes carried on transfer plasmids. Target cells... one copy integrates, selecting by antibiotic-resistance or single cell sorting based on expression of...

...Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O (2014) Crystal...

...simian sarcoma viral (v-sis) oncogene homolog) FLJ12858, PDGF2, SIS, SSV, c-sis PDGFC platelet derived ...PTP, REG RETN resistin ADSF, FIZZ3, MGC126603, MGC126609, RETN1, RSTN, XCP1 RETNLB resistin like beta FIZZ1...tumor necrosis factor receptor superfamily, member 12A CD266, FN14, TWEAKR TNFRSF13B tumor necrosis factor... tumor necrosis factor receptor superfamily, member 17 BCM, BCMA, CD269 TNFRSF18 tumor necrosis factor...tumor necrosis factor receptor superfamily, member 21 BM-018, DR6, MGC31965 TNFRSF25 tumor necrosis factor...tumor necrosis factor receptor superfamily, member 8 CD30, D1S166E, Ki-1 TNFRSF9 tumor necrosis factor...cell differentiation, and apoptosis. Symbol Name Synonyms TNFRSF10B tumor necrosis factor receptor superfamily...

...seed and other stocks of Arabidopsis thaliana and related species. The Arabidopsis Information Resource (...tools for research using plant systems such as Arabidopsis, wheat, maize, rice, tomato, and a variety of... Modular Cloning (JMC) Toolkit Modular cloning chassis and set of functional parts for assembling plant...for creation of fusion constructs in transgenic analysis of plants. Return to top Genome Editing The following... in variety of plant species (e.g. maize and Arabidopsis) and vectors for easy assembly of one or more...Cas9 assembly for fast generation of multiple Arabidopsis mutants. Engineered biocontainable RNA virus ..., 1000000150 2in1 Plasmid Toolkit Plasmids for analysis of protein-protein interactions in planta via ...

...agonist CASP3 Caspase 3, apoptosis-related cysteine peptidase CASP8 Caspase 8, apoptosis-related cysteine peptidase...effects of p53 are in promoting cell cycle arrest, apoptosis, or senescence in damaged cells. The p53 name ...pro-apoptotic effects is especially important to tumorigenesis. Inheriting only one functional copy of the ... 1; also known as GTSE1 BAI-1 Brain-specific angiogenesis inhibitor 1 Bax BCL2-associated X protein Bid...peptidase CASP9 Caspase 9, apoptosis-related cysteine peptidase Cyclin B CCNB1 CCNB2 CCNB3 Cyclin B1, ...ubiquitin protein ligase cytoC Cytochrome c DR5 Tumor necrosis factor receptor superfamily, member 10b E2F-1 ...protein p53 p53AIP1 Tumor protein p53 regulated apoptosis inducing protein 1 p53R2 p53 inducible, ribonucleotide...

...Endo/Exocytosis Superecliptic pHluorin fused to glutamate receptor Glutamate receptor exocytosis and spine...Robert Malinow Endo/Exocytosis Synaptophysin-pHluorin Clathrin-mediated endocytosis is the dominant mechanism...Synaptic vesicle pH sensors for imaging exocytosis and endocytosis (pHuji). Semisynthetic fluorescent pH...pH sensors for imaging exocytosis and endocytosis. Nat Commun. 2017 Nov 10;8(1):1412. Justin Taraska ,... Rise in Free Mg 2+ Ions Released from ATP-Mg Hydrolysis Contributes to Mitotic Chromosome Condensation... FRET sensors to monitor intracellular Zn 2+ homeostasis. Nat Methods. 2009 Oct;6(10):737-40. Maarten ... ATP sensor for Yeast (AT1.03 derived) Dynamic analysis of cytosolic glucose and ATP levels in yeast using...

...promotes protein and lipid synthesis, and increases energy production through glycolysis to fuel these energy-intensive...key metabolic regulator, correlates with poor prognosis in many cancers. Cancer...Ras Pathway p53 Pathway Tackling Cancers’ Drug Resistance with a New Screening Kit Background mTOR Pathway...catabolic processes including autophagy and lysosome biogenesis. mTORC2 does not respond to nutrients, but it... either mTORC1 or mTORC2 correlates with poor prognosis in many cancers, including breast, colon, ovarian...protein S6 kinase B1 TSC1/2 TSC1 TSC2 Tuberous sclerosis VHL Von Hippel-Lindau tumor suppressor Return ... Kit (Sabatini & Wood) Tackling Cancers’ Drug Resistance with a New Screening Kit (from David Sabatini...

...( nicks ). ID Name Insert Function Antibiotic Resistance Activity PI 52962 lentiCas9-Blast Sp Cas9 Cut...codon-optimized S. pyogenes Cas9 protein and blasticidin resistance from EFS promoter. Lentiviral backbone. Zhang...transcriptional activator . ID Name Insert Antibiotic Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A,...lenti vector encoding dCAS9-VP64 with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE...to combinatorially index cells for single-cell analysis of clonal dynamics. This library expresses EGFP...

...including resistances to chemotherapy drugs and toxins, cell viability, and tumor metastasis. One of the...editing systems include targeted AID-mediated mutagenesis (TAM) and CRISPR-X , in which Cas9 is fused to...nsertion of T ransposable E lements by G uide R NA- A ssisted T argeting), developed by Samuel H. Sternberg’s...large scale edits. PASSIGE ( P rime- E diting- A ssisted S ite- S pecific I ntegrase G ene E diting), developed...desired phenotype. The ease of gRNA design and synthesis, as well as the ability to target almost any genomic...simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single transcriptional...than other activators/repressors due to their persistence and heritability. Not all modifications are stable...

...at Addgene. Amyotrophic Lateral Sclerosis Amyotrophic lateral sclerosis (ALS), sometimes referred to as...with siRNA-resistance SigmaR1 constructs . (Sawyer et al. Dev Cell. 2024) Image exocytosis with VAMP2/... ALS, and Huntington’s Disease. These diseases persist for many years and lead to a major burden on the...or way to prevent progression of PD. Treatment consists mainly of managing symptoms through medication...

...Rinehart Lab Phosphoprotein Reagents Phosphoprotein Synthesis Reagents References & Protocols Additional Resources... ). The phosphorylation sites (“phosphosites”) consist of a central phosphoserine site flanked by 15 amino...ambiguity. Rinehart Lab Improved Phosphoprotein Synthesis Reagents Type ID Item Strain 192872 rEcoli XpS... Mode #1 Library Plasmid 69118 E17TAG GFP zeo resistance Plasmid 68307 SupD Strain 68306 C321.ΔA.Δserb.Amp...recombinant phosphoproteins using cell-free protein synthesis . Nature Communications , 6 (1). https://doi.org...phosphoserine insertion during Escherichia coli protein synthesis via partial UAG codon reassignment and release...

...226567 siRNA-resistant SigmaR1-mCherry SIGMAR1 mCherry CMV ALS Gia Voeltz 226569 siRNA-resistant pLV-SigmaR1...Alzheimer's Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Frontotemporal dementia (FTD), Huntington's...Zena Werb 19228 pBS ALS2 ALS2 T7 Primary lateral sclerosis juvenile Cliff Tabin 19457 pcDNA5/FRT/TO HSPA1L...Parkinson's, Kufor-Rakeb syndrome, Neuronal ceroid lipofuscinosis Veerle Baekelandt 171771 miR3 Mm ATP13A2 ATP13A2...Parkinson's, Kufor-Rakeb syndrome, Neuronal ceroid lipofuscinosis Veerle Baekelandt 171786 MSCV-Fbxo7-IRES-GFP...Parkinson's, Kufor-Rakeb syndrome, Neuronal ceroid lipofuscinosis Veerle Baekelandt 171821 pLenti HsATP13A2 D962N...Parkinson's, Kufor-Rakeb syndrome, Neuronal ceroid lipofuscinosis Veerle Baekelandt 171822 miR5 Mm ATP13A2 ATP13A2...

...transcriptional activation of endogenous genes. It consists of three components: A nucleolytically inactive...lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding the...HSF1 activator helper complex with a 2A Hygro resistance marker 61427 : lentiviral sgRNA cloning backbone...loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker. Contains BsmBI sites for insertion of...enables Cre/loxP recombination and fluorescence assisted sorting of cells and nuclei in addition to sgRNA...facilitates Cre/loxP recombination and fluorescence assisted sorting of cells and nuclei in addition to sgRNA...

...developmental biology. With its rapid development (embryogenesis is complete in ~3 days), short generation time...Keith Joung Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9...zebrafish - Bret Pearson Lab MAZe: a tool for mosaic analysis of gene function in zebrafish - Julian Lewis Lab... loss and ablation of previously resistant cell types. CreLite - George Eisenhoffer Lab. ...Neurodegenerative Disorders - Nicholas Cole Lab. A kit consisting of plasmids with four promoters, six fluorophores...

...RCIscript-GoldyTALEN is designed for in vitro synthesis of TALEN mRNAs. Both 5’ and 3’ Xenopus β-globin...custom repress gene expression in yeast. TALORs consist of the DNA-binding domain of a TALE, along with... promoter or can be achieved by in vitro mRNA synthesis from the T7 promoter. Truncations were introduced...pCAGGS-TAL-NC2, are mammalian expression/mRNA synthesis vectors containing unique shorter N- and C-terminal... has successfully been employed for deletion mutagenesis and gene targeting in human and mouse pluripotent...

...CRISPR plasmid kits. CRISPR Software Sanger Indel Analysis ICE (Inference of CRISPR Edits) This open access... opens in a new window) . Deep Sequencing Data Analysis CRISPResso CRISPResso2 is a robust and easy-to-use...easy-to-use computational pipeline for the analysis of genome editing experiments. It enables rapid and intuitive...Link opens in a new window) . MAGeCK Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout (MAGeCK). ...zebrafish, fly, worm, Japanese rice fish, maize, Arabidopsis, and rice. Proof-of-concept performed in zebrafish...

...stabilization or even limited recovery of skills occurs. Diagnosis of Rett syndrome is currently based on specific...window) HRV Metrics Toolbox - algorithms for data analysis to calculate heart rate variability (Link opens... PMID: 11809720 Tarquinio et al. 2015. Age of diagnosis in Rett syndrome: patterns of recognition among...among diagnosticians and risk factors for late diagnosis. Pediatr Neurol . 52, 585-591.e2. PMID: 25801175...25801175 Tillotson and Bird. 2019. The Molecular Basis of MeCP2 Function in the Brain. J Mol Biol . 17;S0022...

...attR2); Zea mays Ubi1 promoter and Hygromiycin resistance marker Also see more empty backbones on our Plant...with N-terminal Myc tag and attB for Drosophila transgenesis His Epitope tag pEZYmyc-His - C-terminal ...transformation plasmids with G418 selection Find nematode transgenesis plasmids with Neomycin selection Puromycin Mammalian...Variant of lentiCRISPR v2 that confers hygromycin resistance pRGEB32 - Express sgRNA/PTG with rice snoRNA ...loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker pLenti CMV/TO GFP-Zeo DEST (719-1) - 3rd...

...of COVID-19 and related coronaviruses. Addgene assisted this effort by providing a platform for the distribution... a cysteine endosomal protease that triggers proteolysis of SARS-CoV Spike protein, activating its membrane...Spike Receptor Binding Domain Site-Saturation Mutagenesis Libraries Screening Jesse Bloom Tyler Starr Libraries...and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe...

..., inflammation, metabolism, fibrosis, atherosclerosis, and carcinogenesis. The defining feature of the...

...Toxoplasma (toxoplasmosis), Trypanosoma (African sleeping sickness), Cryptosporidium (cryptosporidiosis), Leishmania...Leishmania (leishmaniasis), and others have been the focus of biomedical research for decades. Recent developments...

...Caenorhabditis elegans. Mos1 Based Transgenesis miniMos Transgenesis Kit - Erik Jorgensen Lab. This kit...transparent body of the C. elegans hermaphrodite consists of 959 somatic cells, which makes them attractive...