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...facilitation, DNA amplification, and high-quality quality control measures such as titering, purity assessment, sterility...is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity, is easy ...serotype? Learn more about AAV serotypes and tissue tropism on Addgene's blog . All-in-One Pricing The price...centrifugation purification Addgene’s comprehensive quality control process: Titering Purity assessment Sterility testing...produce the prep, and perform comprehensive quality control processes to make sure it is ready for your experiments...your AAV vector experience, from methods to troubleshooting insights, you make research more reproducible...gap between published methods and real-world troubleshooting. Eligible Plasmids The following plasmids are...

...81070 rAAV2-retro helper AAV retrograde AAV packaging plasmid, expressing Rep2 and retrograde capsid Alla... transgene or genetic elements of interest to introduce in cells. A packaging plasmid (or Rep/Cap plasmid...non-standard), expressing Rep2 and PHP.eB Cap controlled by tTA-TRE amplification system Viviana Gradinaru...(non-standard), expressing Rep2 and PHP.S Cap controlled by tTA-TRE amplification system Viviana Gradinaru...non-standard), expressing Rep2 and PHP.V1 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...plasmid (non-standard), expressing Rep2 and B10 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...plasmid (non-standard), expressing Rep2 and B22 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...

... undergo stringent quality control by Addgene or a trusted quality control partner before distribution...are initially titered by spectrophotometry using a NanoDrop spectrophotometer. If needed, the sample is...Addgene's antibody production methods and quality control processes for purified recombinant antibodies. ...production protocols, titering methods, and quality control are described below. Information for a specific...samples with >90% purity are distributed. Quality Control Each lot of antibody undergoes testing, either ...fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections are...Protein expression is compared to an untransfected control and the protein size is confirmed. Residual Plasmid...

...visualization, study of protein interactions, controlled gene expression, reporter plasmids and genome...Purification Protein Visualization Protein Interactions Controlled Expression Reporter Plasmids Genome Engineering...arrays of zinc finger nucleases. Addgene Blog Introduction to FRET Bimolecular Fluorescence Complementation...Studying Protein–Protein Interactions Return to top Controlled Expression It isn’t always advantageous to constitutively...stationary phase. In these cases, you may want to control protein levels or to turn on expression only at...contains plasmids whose expression levels can be controlled by a variety of small molecules, light, and temperature...Escherichia coli Yingfeng An Additional Addgene Controlled Expression Resources Check out our Tetracycline...

...systems, how to use them to modify the genome or control gene expression, and find plasmids for your experiments...engineering tools used to precisely manipulate DNA and control the expression of specific genes in cells or organisms...recombinases can be used to modify the genome or control gene expression or skip ahead to browse highlighted...Summary of Cre, Dre, and Flp Recognition Sites. Controlling Recombinase Expression By expressing the enzyme...promoters or inducible systems, you can precisely control and optimize expression of your gene of interest...fused to a ligand-binding domain such as the human estrogen receptor (ERT2) so that the enzyme is only activated... and C-terminal fragments and placed under the control of different promoters. Expression of both fragments...

...telangiectasia Didier Trono 14581 pSUPER.ATMi ATM H1 Ataxia telangiectasia Didier Trono 14666 pQE9-His-p97...T7 Parkinson's Dmytro Yushchenko 107293 s2. pT7-7-SNCA-5Asp SNCA T7 Parkinson's Dmytro Yushchenko 107294... T7 Parkinson's Dmytro Yushchenko 107295 r. pT7-7-SNCA-5Asn SNCA T7 Parkinson's Dmytro Yushchenko 107296...T7 Parkinson's Dmytro Yushchenko 107297 r-22. pT7-7-SNCA-9Asn SNCA T7 Parkinson's Dmytro Yushchenko 107425...neuroaxonal dystrophy Michael Ward 178167 RNF216_Halo_C_allele RNF216 Halo Hypogonadotropic hypogonadism...Ataxia Pietro De Camilli 194005 VPS13D1-1576-(Y1253S) VPS13D GFP CMV Spinocerebellar Ataxia Pietro De Camilli...Parkinsonism Pietro De Camilli 232865 mCherry-VAB(VPS13C) VPS13C mCherry CMV Parkinsonism Pietro De Camilli...

...Image from Fenno et al., 2014. INTRSECT INTRSECT (intronic recombinase sites enabling combinatorial targeting...and Flp) recognition sequences within synthetic introns. INTRSECT was first shown as a proof-of-concept...transgenic recombinase-expressing animal lines and retro-grade expressing viruses delivering additional recombinases.... Figure 2: INTRSECT works by inserting short, intronic sequences into the open reading frame (ORF) of...sequences (e.g. Lox sites, FRT sites) inside of the introns (A,D). The addition of these recombinase recognition...recognition sequences enables directional control of the portion of the ORF that is sandwiched between the... combinations of recombinases are present, the introns are excised during mRNA processing, producing a...

...PGKpuro2ABFP - For retroviral delivery of one sgRNA pMKO.1 GFP - Retroviral shRNA expression Retroviral backbones...test the function of your gene of interest in a controlled environment under various conditions. When choosing...tag for mammalian expression (Gateway) pYIC - Bicistronic fluorescent reporter gene with cap-dependent ...expression, see our dedicated Lentiviral Plasmids page Retroviral Easy and safe to use, integrate into host genome...MSCV-IRES-GFP or pMSCV-IRES-mCherry FP - Mammalian retroviral gene expression with GFP or mCherry selectable... luciferase vector for investigating regions controlling transcription See our Luciferase collection of... production AAV-GfaABC1D-MCS--4x6T-WPRE - For astrocyte-selective expression in AAV Dual promoter Separate...

...be measured using fluorescence microscopy or spectroscopy by exciting the donor and detecting the emission...available, we have listed cloning backbones and control plasmids. Donor Acceptor λ Dex QY D λ Aem ε A QY...refer to published reports and include appropriate controls to help interpret your results. See Algar et al...feature plasmids with positive and negative FRET controls or calibration standards consisting of donor and...quantitative two-hybrid FRET Positive and negative controls and cloning backbones for mTurquoise2-mVenus(L68V...towards a quantum yield of 93% Positive and negative controls and cloning backbones for various FRET pairs with...acceptors for mTurquoise2 Positive and negative controls and cloning backbones for red, yellow, and turquoise...

...Ready-to-use AAV available from Addgene's viral service. Control EGFP vectors in various serotypes for serotype ... AAV2, AAV5, AAV6, AAV8, AAV9, AAV11, and AAV retrograde....fluorescent reporters and can be used to compare the tropism of different serotypes. In addition to the standard...list of all 100 µL sized sample, please see our Control AAV Inventory . The catalog ID for the 20 µL sized... catalog item, including the titer and quality control, visit the material page (linked for each catalog...AAV11 37825-AAV11.T 20 µL $ 150 Add to Cart AAV Retrograde 37825-AAVrg.T 20 µL $ 150 Add to Cart pAAV-hSyn-EGFP...

...provide a basic understanding of CRISPR biology, introduce the various applications of CRISPR, and help you...research. CRISPR Basics In this section, we will introduce the basic mechanisms of CRISPR and some of the... nuclease fidelity SpCas9-NG — NG; increased in vitro activity SpG — NGN; increased nuclease activity ...-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages...LmoCas9 and SpCas9. Acr proteins can be used to control both when and where genome editing happens. CASANOVA...Dominik Niopek’s lab , is a way to optogenetically control genome editing by fusing the anti-CRISPR protein...photosensitive LOV2 domain . Several labs have also controlled Acr activity with endogenous cell-type specific...

...CRISPR Experiment Jump To: Plan Your Experiment Introduction Select Genetic Manipulation Design and Clone...generally perform by chemical transfection or electroporation. Expression can be transient, or you can generate... of Cas and gRNA. This method involves using in vitro transcription reactions to generate mature Cas mRNA... to target cells through microinjection or electroporation. The target cells translate the mRNA encoding...you directly combine purified Cas protein and in vitro transcribed gRNA to form a Cas-gRNA complex. You...delivery the RNP to cells using cationic lipids, electroporation, or an endosomal approach. Similar to the mRNA...Weissman, J. S. (2014). Genome-Scale CRISPR-Mediated control of gene repression and activation. Cell , 159 (...

...collections available at Addgene. Amyotrophic Lateral Sclerosis Amyotrophic lateral sclerosis (ALS), sometimes...Reprogram astrocytes to dopaminergic neurons using three transcription factors in a single polycistronic lentiviral...being adopted in the field. Read on for a brief introduction to the genetics associated with some of the ...Symptoms include changes in behavior and emotion, uncontrolled movements, memory deficits, as well as difficulty...neurons in the substantia nigra, one of the movement control centers in the brain. The cause of this neuron ...

...samples of control AAV vectors for trial purposes in serotypes AAV1, 2, 5, 6, 8, 9, 11, and retrograde Retrograde...is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity, is easy .... Addgene’s comprehensive high-quality quality control process includes titering, purity assessment, sterility... Sensors, Glutamate Sensors, Voltage Reporters Controls Fluorescent Proteins, Spagehetti Monster, Brainbow...

...optogenetically controlled Cre/loxP system that enables precise temporal and spatial control of gene expression...horizontal stripes on the sides of the adult body, is a tropical freshwater fish in the minnow family. It has long...refer to Addgene's CRISPR Guide for a general introduction to CRISPR technology or the zebrafish CRISPR...ALS-linked aggregates in real time. Live imaging of astroctyes - Kelly Monk Lab. Live imaging tools to study...study the development and function of zebrafish astrocytes. Knock-in tagging - Michel Bagnat lab. Targeting...

...intracerebroventricular (ICV) delivery in newborns, and retro-orbital (RO) and stereotaxic (STX) delivery in adult...minBG-SYFP2-WPRE3-BGHpA AiP14470 AiE0027m_3xC3 SYFP2 Ventromedial hypothalamic nucleus (VMH) Hypothalamus 230531...minBG-SYFP2-WPRE3-BGHpA AiP12558 AiE0390m_3xC2 SYFP2 Astrocytes Whole Brain 208163 pAAV-AiE0390m_3xC2-minBG-iCre...-iCre(R297T)-BGHpA AiP14801 AiE0390m_3xC2 Cre Astrocytes Whole Brain 208143 pAAV-AiE0390h_3xC2-minBG-SYFP2...minBG-SYFP2-WPRE3-BGHpA AiP13572 AiE0390h_3xC2 SYFP2 Astrocytes Whole Brain 208133 pAAV-AiE0387m-minBglobin-SYFP2...-SYFP2-WPRE3-BGHpA AiP12102 AiE0387m SYFP2 Astrocytes Whole Brain 208166 pAAV-AiE0387m-minBG-iCre(R297T...R297T)-BGHpA AiP13999 AiE0387m Cre Astrocytes Whole Brain 220643 pAAV-AiE2128m-minBG-SYFP2-WPRE3-BGHpA AiP1357...

...expression is driven by the strong CAG promoter or can be achieved by in vitro mRNA synthesis from the T7...promoter. RCIscript-GoldyTALEN is designed for in vitro synthesis of TALEN mRNAs. Both 5’ and 3’ Xenopus...of the DNA-binding domain of a TALE, along with strong nuclear localisation tags and repress transcription... T7 promoter. Truncations were introduced to the N- and C-terminus of the pTAL3 TALEN backbone, which ... included for generating TALEN mRNAs through in vitro transcription as an alternative. Based on transient...

...Production and Quality Control Production Our production process and quality control, with examples of quality...production, purification, and comprehensive quality control. In-Stock Preps 1,000+ preps ready to ship immediately...quality control data. Help Center Articles Additional details about viral vector production at Addgene...various aspects of viral vectors, including troubleshooting advice and explanations of viral vector technologies...

...Optogenetics plasmids encode proteins that are controlled by light. Many of these plasmids encode for microbial...the cell. When using these opsins as a tool to control neuronal activity, scientists generally attach ...localization. Many other types of proteins can also be controlled by light, generally by utilizing a light sensitive...fusions can then be used as an optical switch to control protein interactions, dimerization, localization...the table below for plasmids that use light to control a protein or cellular activity. For example, search...

...viral service encoding tools for monosynpatic retrograde neuronal tracing, to be used with modified rabies...be used with modified rabies virus to perform retrograde neuronal tracing. Note : Addgene does not distribute...pAAV-TREtight-mTagBFP2-B19G AAV1 Ian Wickersham Introduction to Rabies Virus-based Monosynaptic Tracing The...-deleted rabies to specific starter cells, the tropism of this virus was modified so that it could only..., Finke S, Conzelmann KK, Callaway EM. 2007a. Retrograde neuronal tracing with a deletion-mutant rabies...