We narrowed to 731 results for: Sis;
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TypeCollection...is done using the envelope protein of (EnvA) of leukosis virus. EnvA can direct virus infection specifically...
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Nuclear Receptor Signaling Atlas (NURSA) Plasmid Guide: Nuclear Receptors
TypeCollection...receptors play a key role in development and homeostasis, and the receptors and their downstream genes... -
Antibody Production
TypeCollection...applications such as these, the buffer can be removed by dialysis or gel filtration. Titer and Purity Antibodies... -
CRISPR Plasmids - Drosophila
TypeCollection...gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species... -
CRISPR Plasmids - Yeast
TypeCollection...Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species... -
New England Biolabs Cell-Imaging Plasmid Collection
TypeCollection...Peptide bond formation in nonribosomal peptide biosynthesis. Catalytic role of the condensation domain. ... -
Zinc Finger Consortium: Zinc Finger Arrays
TypeCollection...):1471-1501. PubMed PMID 19798082 . Targeted mutagenesis in zebrafish using customized zinc-finger nucleases... -
AAV Packaged on Request
TypeCollection...answer isn’t in the Help Center or you need more assistance with troubleshooting, we want to hear from you... -
CRISPR Plasmids - Bacteria
TypeCollection...Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species... -
TALEN Guide
TypeCollection...available as a kit from Addgene , custom arrays consisting of 12-31 repeats can be assembled and inserted... -
CRISPR Plasmids - Mammalian Expression
TypeCollection...cytosine demethylation by Tet1. These modifications persist over time and are potentially heritable in dividing... -
CRISPR Plasmids - Plants
TypeCollection...U6 BbsI none S. pyogenes Gao 59188 pBlu/gRNA Arabidopsis U6 BbsI none S. pyogenes Stupar 62200 pBUE411... -
Plan Your Experiment
TypeCollection...identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or NHEJ): For large deletions or insertions... -
CRISPR History and Development for Genome Engineering
TypeCollection...DA, Horvath P. 2007. CRISPR provides acquired resistance against viruses in prokaryotes. Science . 315... -
Fluorescent Protein Guide: Subcellular Localization
TypeCollection...cytoskeleton is the cell’s internal framework that assists in sustaining structure and function. The cytoskeleton... -
Genetic Code Expansion
TypeCollection...sufficient levels of Sep in the cytoplasm for protein synthesis. Also see Strain 192872. Jesse Rinehart 69493 ... -
CRISPR Antimicrobials
TypeBlog Post...without a resistance gene, allowing resistant bacteria to multiply, spreading their resistance genes throughout...The crisis of antibiotic resistance is upon us, and the world is unprepared. Each year in the United ...antibiotic resistant bacteria. Even when researchers develop new antibiotics, the onset of resistance is swift...antibiotic resistant bacteria with a weapon that they’ll have less potential to become resistant to? CRISPR...antibiotic resistance Most antibiotics target one of three essential processes: cell wall synthesis, DNA replication...or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria, allowing...propagate their resistance genes. Our society is hoping to combat antibiotic resistance using multiple ... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post...example, we examined ICE analysis of Sanger sequencing data alongside the analysis of NGS-based amplicon ...scalability, the ICE software has two analysis formats: sample by sample analysis, which can compare up to five...synthego.com. Overview of ICE editing analysis Once the analysis is complete, a new screen appears with...also download the entire analysis as a ZIP file by clicking the Download Analysis Data button on the bottom...bottom right of the analysis screen. ICE analysis details Contributions The Contributions tab shows the...tool was initially created to support the CRISPR analysis needs of Synthego’s scientists, and it’s now free...sequencing data and found that the accuracy of ICE analysis results was highly comparable (within r2=0.96)... -
Deep Dive: qPCR
TypeBlog Post... Statistical analysis Getting your RQ value isn’t the end of your analysis process. You’ll still...different statistical analysis methods that pair well with slightly modified CT analysis, so pick one and ...chemistries, primer optimization, controls, and analysis. As a rule of thumb, you can assume that qPCR...chemistry The two most common qPCR chemistries are hydrolysis probes (which you probably know as TaqMan) and...multiplexed Single amplicon validation technique Hydrolysis Probes (Taqman) Low; can be reduced with a ... Yes, one Yes Yes Agarose gel or uMelt analysis dsDNA dye (Sybr) High No No No ... Yes, two Yes Yes Agarose gel or uMelt analysis Hairpin probes Low Yes, one Yes Yes... -
New Tool for Lineage Tracing: The ClonTracer Library
TypeBlog Post...cancer development, metastasis, and drug resistance, Bhang says “the [drug] resistance story panned out ...cells that represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates...found that similar barcodes consistently made up the majority of the resistant populations. In fact, 88%...apply [it to].” Using ClonTracer to study drug resistance Although Bhang and co-workers started using the...determine whether or not cancer cell populations resistant to drug treatments arise during treatment or are... outcomes (see schematic depiction below): 1. Resistant cells among the separate replicates would contain...abundance before and after a treatment. The library consists of a pool of lentiviral vectors with roughly 73... -
Five Popular Model Organisms, Part 2
TypeBlog Post...is as a biomedical model organism. Arabidopsis thaliana Arabidopsis thaliana is one of, if not the most...manipulation and mutagenesis techniques, are available to the scientific community. The Arabidopsis genome project...beginning of widespread Arabidopsis genetics research development. Arabidopsis thaliana was fully sequenced...for use in CRISPR and Tol2 transposase induced transgenesis. Xenopus Xenopus, or frogs, are one of the ...understand human disease alleles during and after embryogenesis. Addgene has over 600 plasmids that contain ...characterized genome, and widespread utilization, Arabidopsis has made significant contributions to understanding...understanding plant and crop science alike. Arabidopsis has many of the traits that make an ideal model organism... -
Plasmids 101: Shuttle Vectors
TypeBlog Post...have a plan to ensure it persists in both contexts. For a vector to persist in... You need... Bacteria... an antibiotic resistance gene to allow for antibiotic selection, but these resistance systems don’t always... antibiotics also block protein synthesis, and the neomycin resistance gene protects against any of them...single lecture slide. That shared language is the basis of shuttle vectors, which can function in unrelated...like penicillin and ampicillin inhibit cell wall synthesis. Eukaryotes, with no cell walls, can grow alongside...eukaryotes. For example, puromycin inhibits protein synthesis across kingdoms by targeting a shared aspect of... of ribosomal function. A puromycin resistance gene on your vector can protect both plasmid-carrying bacteria... -
Plasmids 101: Knockout/Knock-In Plasmids
TypeBlog Post... designed to insert a resistance gene. The vector contains a neomycin resistance gene (NeoR) flanked by...insert and later remove a resistance gene. In this construct, the neomycin resistance gene is floxed so that...into the vector on either side of an antibiotic resistance gene. To avoid selecting for cells in which the...perform a positive selection for the antibiotic resistance gene and then a counter selection for the negative...removed from the chromosome and and replaced by the resistance gene.The gene is thus disrupted, or knocked out...occurred. Neomycin, puromycin, and hygromycin resistance genes are commonly used for positive selection...positive selection marker. Since an antibiotic resistance gene can affect a cell's phenotype, researchers... -
How to Keep a Lab Notebook for Bioinformatic Analyses
TypeBlog Post...well as you perform analysis after analysis after analysis… Treat every analysis like a wet lab experiment... my thesis project a few years ago when my advisor and I decided to do a massive RNA-seq analysis. I had...you to perform this analysis. Having a clearly stated, specific goal for each analysis will help you locate...Before you begin, briefly state the goal of the analysis, including background on other analyses that have...approach that you will take to help you plan your analysis. There is no need to go into a lot of detail –...Always, always, write a brief conclusion of your analysis, even if your conclusion is “this approach is ...ideal because…” Including a conclusion for each analysis or task will keep you from repeating your work... -
The Breast Cancer Microenvironment: A Tumor’s Backstage Team
TypeBlog Post...of? A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The microenvironment...altered fibroblasts and promote the development and metastasis of tumors through inciting inflammation. Speaking...microenvironments affect tumor spread and drug resistance? Although we’ve seen how multiple types of cells...story. It’s essential for a tumor to spread and to resist drugs. And to do that, let’s take a look at exosomes... case, exosomes may transport miRNA from drug-resistant cells to sensitive ones. Though this process is... expression of TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells...cells proliferate and cannot undergo apoptosis, thus multiplying uncontrollably and building up tumors.... -
Modulate the Activity of 17 Signaling Pathways with One Kit!
TypeBlog Post...involved in drug resistance Wood wanted to find out which particular pathways drive resistance to specific ...relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said...comprehensive understanding of how cancers become resistant, an emerging theme is that they do so by activating...proliferation, survival, differentiation, and apoptosis. For each of these 17 pathways, they identified... a time, you could identify those that confer resistance to drugs,” in a rapid, straightforward, and scalable... for use in screening for pathways that drive resistance to drugs in vitro, and they’ve shown that it ...confer any phenotype of interest, not just drug resistance,” he said. “It could [be used to screen for] ... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab...mini-Tn5 vector that can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters...this vector, called pBAM1, by conducting random mutagenesis in the soil bacterium Pseudomonas putida and ...constitutively expressed, reports on the balance of biogenesis and mitochondrial degradation in vitro and in...the complexity of assembling recombination of transgenesis constructs? A new cloning system combining the...maximum of 24 elements assembled into a final transgenesis construct. Plasmids constructed using Golden...recombination template vectors, to perform multiple site mutagenesis and create complex fusion or recombination vectors... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...from Addgene consists of two plasmids: pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries... targeting the gene of interest and kanamycin resistance E. coli carrying the phage recombineering machinery...recombineering is unsuccessful and the wild-type sequence persists, Cas9 will cleave the gene of interest, inducing...Insertion of Transposable Elements by Guide RNA-Assisted Targeting) and the Zhang lab's system called CAST... enable gRNA-directed transposition. INTEGRATE consists of four major components including (1) a CRISPR...Streptococcus pneumoniae. Robert Husson lab's M. tuberculosis CRISPRi plasmids A group from the Husson lab... used CRISPRi to study essential genes in M. tuberculosis. dCas9 and a gRNA are expressed on pRH2502 and... -
Plasmids 101: Dimers and Multimers
TypeBlog Post...of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique that can detect multimers...Levene SD. Analysis of DNA topoisomers, knots, and catenanes by agarose gel electrophoresis. Methods Mol...stability study and topology analysis of plasmid DNA by capillary gel electrophoresis. BioProcess Int. 2006 ...the supercoiled plasmid monomer, and at <6 kb, consistent with the size of a supercoiled plasmid dimer.... multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene therapy...Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med. 2020;20(10):798-805. doi: 10.2174... He Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence of plasmid ... -
Plasmids 101: The Wide World of Natural Plasmids
TypeBlog Post... Blog Plasmids 101: Antibiotic Resistance Genes Antibiotic Resistance: An Old Solution but a New Problem...environments. The genes encoding these functions form the basis of many molecular biology tools! Figure 1...natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense Genes allowing...its host. For example, cells with a penicillin resistance plasmid can grow in spite of a hostile Penicillium...fungus trying to kill them off. While antibiotic resistance-mediated selection is indispensable for cloning...cloning work, antibiotics and plasmid-encoded resistance to them are the tools of a millennia-old microbial...like colicin typically also carry the necessary resistance genes to help cells survive their own secretions... -
Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More
TypeBlog Post..., making them ideal for studying vertebrate embryogenesis. To illustrate some of the reasons that zebrafish...in developing embryos are available. Additional analysis tools have been developed to help parse and visualize...1990’s, two revolutionary large-scale random mutagenesis screens produced ~1,500 zebrafish mutants that...to study development and disease. Additional mutagenesis projects have since been initiated and today ...gene deactivation tools for zebrafish Random mutagenesis screens provided a major toehold into understanding...the zebrafish is an ideal system to observe embryogenesis live. The origins, divisions, and movements ...tracing defined a high-resolution map of early embryogenesis in time and space, and was also used to precisely... -
Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names
TypeBlog Post...many contexts within the fly including oogenesis and cytokinesis. Why name this protein after a vegetable...hedgehog exhibit abnormal segmentation during embryogenesis. Typically, regularly spaced denticles (bristles...such as cell movement/motility, development, cytokinesis, and muscle contractions (think skeletal muscle...monster, a meiotic arrest gene required for spermatogenesis One of our smaller conference rooms is named... meiotic arrest gene that is necessary for spermatogenesis. Flies that lack cookie monster are sterile...to a failure of primary spermatocytes to enter meiosis. Testis of cookie monster mutant flies are filled...White-Cooper, 2003). Why call a gene required for spermatogenesis after a Sesame Street character? The scientists... -
RUBY-Red Siliques
TypeBlog Post...antibiotic/herbicide resistance and fluorescent proteins. Selection using resistance genes usually involves... editing, transformation may be identified by resistance to an antibiotic. Successful editing of transformants...occurs in T0 plants (from callus) or T1 plants (Arabidopsis floral dip). In the next generation the CRISPR...planning to use it to simplify my work with an Arabidopsis mutant that can’t set seeds. I study the development...and a spontaneous step. (B) RUBY is a reporter consisting of these three enzymes expressed under the same... Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following floral dip transformation next...without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing seeds from ... -
CRISPR 101: Homology Directed Repair
TypeBlog Post...of a homologous donor (e.g., sister chromatid, plasmid, etc.), and consists of three pathways: classical...initiates primary synthesis from a template followed by a round of lagging strand synthesis to fill in the...for strand invasion and a primer for DNA repair synthesis. The ssDNA strand then displaces one strand of...classical double-strand break repair (DSBR), synthesis-dependent strand-annealing (SDSA), and break-induced ... template to serve as a primer for DNA repair synthesis, ultimately leading to the formation of double...manner, this results in a non-crossover event. Synthesis-dependent strand-annealing (SDSA) pathway SDSA... chromosomal aberrations. Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination... -
Plasmids 101: Common Lab E. coli Strains
TypeBlog Post...eliminating stringent factor Allows RNA synthesis in absence of protein synthesis Ptrc-ccdA Propagation of ccdB-containing...of 30S ribosome Confers resistance to streptomycin Tn10 Confers resistance to tetracycline Additionally...uptake, iron uptake receptor mutation. T1/T5 Phage resistance gal Mutation in galactose metabolism pathway...only gyrA, gyrA96 DNA gyrase mutation Confers resistance to nalidixic acid hsdRMS hsdR(rk-, mk+) Unmethylated...constitutive expression of genes for deoxyribose synthesis Allows uptake of large plasmids hee High electroporation...source for growth proAB mutation in proline biosynthesis pathway Requires exogenous proline source for...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R ... -
The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It
TypeBlog Post...the forefront in the so-called “reproducibility crisis.” Results in preclinical, biomedical and psychological...be able to look under the hood at the raw data, analysis code, and other materials. The use of reporting...collected, what the hypotheses are, and how each hypothesis will be tested with a specific statistical test...Preregistration makes clear the distinction between hypothesis testing work, in which a-priori hypotheses are...are tested with a new experiment; and hypothesis generation, in which data are investigated to look for...unexpected trends or differences between groups. Hypothesis generation is critical for advancement, but we...too often encouraged to present that work using hypothesis testing statistical tools that generate a p-value... -
27 Hot Plasmids from 2016
TypeBlog Post...including but not limited to cell growth and drug resistance. Recently, the Wu Lab at Texas Tech developed...Virus (WNV) induced cell death. The Wu lab library consists of 77,406 individual gRNAs targeting a total of...approved drugs against the entire kit; a parallel analysis that successfully identified new, highly specific...alone involves much literature searching, DNA synthesis, and plasmid assembly. Luckily, researchers from...Moduclar Cloning (CIDAR MoClo) Parts Kit. This Kit consists of a variety of plasmids containing many different...processes such as differentiation, proliferation, and apoptosis. In humans, mutations that permanently activate...plasmids to the scientific community. This collection consists of plasmids containing genes from a crowd-sourced... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog Post...mechanisms of the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer have been... process analogous to the development of drug resistance in bacteria, the tumor cells that escape the ...is a region of incessant chemical activity, and consists of other cell types, especially immune cells and... promotes the production of VEGF to increase angiogenesis around the tumor. This in turn regulates the... tumor cells spreading to other organs during metastasis. Evading Recognition Tumor cells over-express...damage healthy cells, and often cannot eradicate metastasis or eliminate the probability of a relapse. Chemical...the immune cells that aid tumor formation and metastasis, block the cell-surface proteins that help escape... -
Mesothelioma - Causes, Symptoms, and Treatment
TypeBlog Post...time from first asbestos exposure to the time of diagnosis) is about 35 years, but it usually ranges from...Stage 4 cancer. These later stages of a cancer diagnosis indicate, for Stage 3, that the cancer is large...originated to another organ in the body. Such misdiagnosis and late discovery mean that most mesothelioma...mesothelioma patients have an extremely poor prognosis. Together, the cause, long latency period, and rarity of...options: Early detection, immunotherapy, and angiogenesis inhibitors New research is also going into two...the best ways to improve the traditionally dire prognosis for mesothelioma patients is to catch the disease...mesothelioma much earlier, thus improving the patient’s prognosis significantly, but also to monitor those who may... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog Post...for the analysis. (C) A more liberal gate allows more cells to be considered for the analysis. Depending...duplicate events, and the Live/Dead gate to focus your analysis on what’s biologically active. After that, the... of tissue processing prior to running my flow analysis, I can however clearly identify my lymphocyte ...good to ask around for advice before running your analysis. In this case, I and other lab members had worked...acquisition voltage (not shown) used in my flow analysis I can spot my lymphocyte population distinct from...beginning should not have a great impact on your analysis, and I often choose a looser gate here. I am not...worried about including irrelevant events in my analysis, as these will get excluded through further, more...