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...Parasite Resources Fully functional Cas enzymes will introduce a double-strand break (DSB) at a specific location...frameshifts, creating loss of function alleles. To introduce specific genomic changes, researchers use DNA ... tables below for CRISPR plasmids designed to introduce a DSB. To learn more about NHEJ, HDR, and other...

...find current pricing information Standard Quality Control Plasmid verified by next generation sequencing ...Plasmid Description Industry PI FAQ What Quality Control analyses are performed on my cloning grade DNA ... and 260/280 ratio has been determined by spectrophotometer. Other methods of quantification may yield...

...blocks the progression of the RNA polymerase, thus strongly decreasing the expression level. Mismatches between...target sequence lower the repression level. By controlling the number of mismatches we can obtain a range...reporters, mCherry and sfgfp in this case, can be controlled using a plasmid‐borne CRISPR array coding for...

...plasmids are in a lentiviral backbone, with some retroviral and AAV backbone options. Plasmids for constitutive...protocols Lentiviral packaging plasmids Web Resources Trono Lab Lentivectors Toolbox Optimized shRNA design ... in a new window) . Wiznerowicz, M., Szulc, J., Trono, D. (2006). Tuning silence: conditional systems ...

... packaging capacity, high efficiency in driving strong transient gene expression in both dividing and ...pShuttle-CMV-F2A-T2A-Venus For production of viruses with multicistronic expression and a Venus tag Wang 50959 ShuttleMCP...-CMV-F2A-T2A For production of viruses with multicistronic expression Wang Return to top Adenoviral Plasmids...

...HIV-based lentiviral particles Murine leukemia-based retroviral particles Vesicular stomatitis virus particles...Description Mutations Industry PI Due to export control regulations, MERS and SARS-CoV materials are available...Lentiviral Envelope and Packaging Plasmids Popular γ-Retrovirus Envelope and Packaging Plasmids Reporter Plasmids...

...apamycin (mTOR) is a key metabolic regulator controlling cell growth and proliferation. mTOR, a serine...in cancer promotes the conditions needed for uncontrolled cell growth and proliferation, including an ...Kris Wood) References mTOR signaling in growth control and disease. Laplante M, Sabatini DM. Cell. 2012...

...CRISPR Read Addgene's CRISPR Guide for a general introduction to CRISPR technology. Addgene's worm CRISPR ...silencing in the germline. A "FLP-Out" system for controlled gene expression in Caenorhabditis elegans. - ...Hubbard Lab. Plasmids for the temporal and spatial control of gene expression in C. elegans by combining expression...

... CRISPR/Cas9 constructs using a high efficiency intron-optimized SpCas9-coding gene in plants. CRISPR ...Kate Creasey Krainer , Vladimir Nekrasov , Nicola Patron Additional Genome Editing Resources Check our Plant... and experimental resources that relate to the control of gene expression in the grasses. The TapRoot ...

...the location and type of MECP2 mutation can be a strong predictor of symptom severity. Broadly, early gene... in a new window) RSRT iPSC lines and isogenic controls , patient-specific Infinity BiologiX - RSRT cell...refer to Addgene's CRISPR Guide for a general introduction to CRISPR technology or the mammalian CRISPR...

...application-specific validation and consistent quality control measures. We work with trusted partners and experts... by sharing our protocols and quality control methods. Learn More Guide New to using antibodies...

...Drosophila Plant Yeast Resources Cas nickase mutants introduce gRNA-targeted single-strand breaks in DNA instead...mutants can also be used with a repair template to introduce specific edits via homology-directed repair (HDR...

... Looking for non-targeting control vectors? Search our catalog for control gRNAs . All-in-one Plasmids...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...Acknowledgement (RSA). This form can be completed electronically by the scientist who will be using the materials...

...stress granules 114411 TNNI1-mEGFP AICSDP-34 mEGFP Troponin I, slow skeletal type Sarcomeric thin filaments...Nuclear speckles 159744 DMD-mEGFP AICSDP-55 mEGFP Dystrophin Costameres 159745 TFAM-mEGFP AICSDP-72 mEGFP ...

... These AAV encode tetracycline-inducible tools/controls and tetracycline transactivators that can be used...TEVp EF1a-driven, Cre-dependent Cre-dependent, bicistronic expression of designer pro-taCasp3 and TEVp for...

...construction of RAS pathway gene expression vectors for in vitro and in vivo research. The RAS Initiative - at the...colorectal cancer. Zenonos K, Kyprianou K. World J Gastrointest Oncol. 2013 May 15; 5(5): 97–101. doi: 10.4251...

...fluorescent protein) in striatal Pvalb+ neurons. Note strong cortical Pvalb and L5 ET neuron expression. Jonathan...3xCore version of DLX enhancer). AAV vector for strong & rapid SYFP2 expression in forebrain GABAergic...

...pHIV-Zsgreen - Mammalian Expression (this is a bicistronic vector rather than a fusion protein vector) ZsGreen1...- Mammalian Expression pMXs-mRFP1 - Mammalian Retroviral Expression mCherry 587 610 16 4.5 15 min Monomer...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

...Biofuels Natural Product Pathways Efflux Pumps Flux Control Metabolism Plasmids Search the table by keyword...

...treatment of neuroendocrine cancers. As such, NETRF strongly encourages its funded scientists to make their...

...Optogenetics Plasmids Use light to activate, inhibit, and control molecular pathways or cells. Chemogenetics Guide...

...construction of RAS pathway gene expression vectors for in vitro and in vivo research. TP53 Mutagenesis Library :...

...Bacterial Expression Plasmids (empty backbones/controls) ID Plasmid GenBank Key Features 26092 p15TV-L...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

...Plasmids Lentivirus Adeno-associated Virus (AAV) γ-Retrovirus Adenovirus All Viral Plasmids Curated Topics ...

...specific small molecules, allowing a scientist to control signaling within cells expressing these engineered...

...for the AAV1, AAV5, AAV6, AAV8, AAV11, or AAV Retrograde serotypes and 2 x 10 12 GC/mL for the AAV2 serotype...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

...enzymes and related tools are available: PE2 — introduction of five mutations in RT enzyme PE3 — PE2 plus...

...HaloTag (Link opens in a new window) technology puts control of the function and properties of a fusion protein...

...vectors to express your gene of interest under the control of a variety of promoters. Professor Oka and the...

...new technology, and we hope to continue to have a strong scientific partnership with both the labs that ...

...traffic, they also play critical roles at cilia, centrosomes, the plasma membrane, lipid droplets, the nucleus...

... Plasmid Description Industry PI Due to export control regulations, MERS and SARS-CoV materials are available...

...Yannick Doyon's lab deposited a plasmid which introduces an N- or C- terminal affinity tag (3xFLAG-2xSTREP...

...undergo application-specific validation and quality control by Addgene as well as by our trusted partner labs...

...specificity, or retrograde transport in neurons. (b) Genetic cargo can be customized to control transgene expression... Capsid Published Tropism Model Organism Mouse Strain-Specific Tropism?  Reference AAV-...and astrocytes Mice No (LY6C1)   Hanlon et al., 2019 AAV-BI-28 CNS: neurons, astrocytes, oligodendrocytes...-Specific Tropism?" column. For a complete list of mice strains with LY6A-dependent tropism, see S3 table...derived from AAV9 using iTransduce, shows tropism toward astrocytes and neurons, with greatly increased efficacy... appears to show a broader tropism with a potential bias toward astrocytes (Chuapoco et al., 2023). Additionally...systemic AAVs with endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications...

...developed a new AAV variant, rAAV2-retro, that migrates retrogradely at higher rates than any other AAV... AAV2-retro vectors with a variety of transgene/promoter combinations. Check out our retrograde AAV blog...requires both antero- and retrograde labelling, synaptic restriction as well as controlled monosynaptic and polysynaptic...Selective retrograde transsynaptic transfer of a protein, tetanus toxin, subsequent to its retrograde axonal...bodies to the terminals of their axons; while retrograde tracing outlines neurons in the opposite direction...axons to their cell bodies. Anterograde and retrograde tracing take advantage of existing transport ...myosin, and enzymes for transmitter synthesis. Retrograde transport is used for transport of endocytosed...

... What kind of control? On, off, tunable, and/or inducible? How long do you need control? Will temporary...temporary control (a few days) be enough, or do you want a permanent control mechanism? Where is the control...transcription instead of introducing a DNA double strand break. Transient introduction of CRISPRi or inducible...vector! On/off control all the time If your experimental system requires tunable control - or maybe you...price of tunable control These systems can be easily implemented for exogenously introduced proteins, as ...prior to introduction of the construct into cells. However, if you would like tunable control of an endogenously...forever? You don’t need to resort to the force to control protein expression in mammalian cells: easy to ...

...viruses: gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus. Gamma-retrovirus Retroviruses...type of gamma-retrovirus is Murine Leukemia virus (MLV). Naturally isolated gamma-retroviruses have been ...Lentivirus is a type of retrovirus and shares many common features with gamma-retrovirus including genome architecture... nucleus as gamma-retroviruses do. Fun fact: Since lentivirus and gamma-retrovirus are so closely related...expression (Robinson, 1982). To add insult to injury, retroviruses generally have moderate to high immunogenicity...immune response in the host. So why use a gamma-retrovirus in the lab? Well, they are essentially evolutionarily... types with a high efficiency. However, gamma-retrovirus infection is restricted to dividing cells only...

...A common means to achieve ligand control is to fuse a mutant estrogen receptor (ER) ligand binding domain...PSD95-mcherry) under the control of Cre recombinase. After in-utero electroporation, this plasmid mix results...tracking for instance) can be controlled with FLP-FRT. For further temporal control, Tamoxifen inducible Cre...system, but it only gives you a single level of control without modification - either Cre is there or it...for site specific (and often cell type specific) control of DNA recombination and gene expression can be...addition, a variety of means to spatiotemporally control FLP and Cre expression have been developed. Read...FLP and Cre enable additional levels of genetic control. What is FLP FRT and how does it work? FLP recombinase...

...Include proper controls To ensure accurate data analysis, include a complete set of controls. Ideally, every...will have a negative control that does not express the target and a positive control that does. The negative...of experiments easier and faster to perform. Introduction to flow cytometry Flow cytometry allows users... (3): An optics system A fluidics system An electronics system  Cells are first resuspended in a pressurized...fluorophore or fluorochrome-stained protein will emit a stronger photon than a cell expressing low levels of the... further the shift of a cell from the negative control, the higher the level of expression. A routine ...triton-x, saponin, or tween. Take care when using strong detergents like triton-x and tween as extended ...

...larger amount of vector using a serotype with strong CNS tropism (4). If axonal transport is not desirable...receptor physiology and the effects of metabotropic and ionotropic influences on overall cell function. ... thanks to our guest blogger Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student at ...post was contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University...known is that different serotypes have different tropisms but, as you can see, AAVs of each serotype can... asking questions!  Table 1: AAV Serotypes and Tropism Experiment  Detection Method CNS AAV1, AAV2...>AAV8>AAV6>AAV5>AAV1>AAV2 This preferential tropism for different brain regions is largely due the ...

...Moloney murine leukemia virus (MMLV)-derived retroviruses. Retroviral vectors, such as pMXs or pMSCV, infect...delivery methods. Retrovirus has a cloning capacity of ~8 kb, so either a single polycistronic vector or multiple...10 kb) than other retroviral vectors. This makes it possible to use a polycistronic cassette that expresses...Mechanism Efficiency Safety Pros Cons MMLV-derived Retrovirus Integrating High Low Efficient and Stable Genome...Need to sequence cells to verify excision didn’t introduce mutations Adenovirus Non-integrating Low Medium...required for translational use. Viral Vectors Retroviral: Some of the first iPSCs were generated with ... corresponding endogenous pluripotency genes. Retroviral vectors are prone to incomplete silencing of ...

...DNA.  Plasmids are said to be under stringent control when they depend on the host cell’s initiation ...replication independently are said to be under relaxed control, as they only require the host's replication machinery...replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...round of replication. Initiation is the primary control point of DNA replication in bacteria, so the process... the bacterial chromosome and for stringently controlled plasmids. Binding of IHF (green) redistributes...binding is necessary to initiate replication, controlling the amount of active DnaA available is one of...source, ATP binding is one more way for the cell to control DNA replication. A variety of other proteins also...

...target genes. The sequence of the promoter region controls the binding of the RNA polymerase and transcription...promoters is commonly employed in lentiviral and retroviral constructs and we will discuss these in a future... all the time while others are more carefully controlled. Regulated promoters might act only in certain... light. In the cell, promoters themselves are controlled by still other regulatory factors: enhancers,...Additional considerations CMV General expression mRNA Strong mammalian expression promoter from the human cytomegalovirus...some cell types. EF1a General expression mRNA Strong mammalian expression from human elongation factor...promoter hybrids. CAG General expression mRNA Strong hybrid mammalian promoter Constitutive  Contains...

... under the control of the cI857 temperature-sensitive repressor. The temperature-controlled expression...activity in the Co-IP compared of a control Co-IP indicates stronger interaction. The system has been demonstrated...DECEMBER 2016 Restoring yeast prototrophy with plasmids Though auxotrophic markers are used frequently ...complement strains with multiple auxotrophies or using different auxotrophic backgrounds can lead to disparate...their Yeast Prototrophy Kit, a set of plasmids designed to complement unused auxotrophies in Saccharomyces... at a dose and duration sufficient to kill all control cells and surviving “mutant” colonies were deep...GTPases involved in cell signaling pathways that control many cellular processes such as differentiation...