We narrowed to 355 results for: Cre expression vector

... information on viral vectors, including transduction download our Viral Vectors 101 eBook. Bacterial ...can be used to create both transient cell lines, where a gene of interest is expressed but not integrated...bacterial or eukaryotic cells via a virus or viral vector. One example are bacteriophages that attach to ...scientists commonly use phagemids, a DNA cloning vector that contains both bacteriophage and plasmid properties...application of an electrical charge to cells, to increase cell membrane permeability and thus transformation...DNA can be accidentally packaged into the newly created phages. Transfer of this DNA to another cell is...Electroporation on the other hand uses electrical pulses to create transient pores in the cell membrane that genetic...

...CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain GABAergic interneurons...-AAV5 pAAV-CAG-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV8 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...AAV9 pAAV-Ef1a-DIO-PPO-Venus AAV vector for Cre-dependent expression of Venus-tagged parapinopsin driven...combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent viral vectors. Jonathan...AAV-mediated expression of voltage sensor under the Syn promoter, Cre-dependent expression; soma localization...

...transgenic protein for the HEK 293T cells to express: CD45. Upon expression, CD45 is incorporated into the plasma...CD45, but is separately expressed. (The reason for having a separately expressed transduction reporter ...manage to express CD45. Thus, if transduction efficiency is much higher than CD45 expression, this could... example, point at issues with the double-expression vector.) In our flow experiment, we'll want to identify...CD45+ cells (CD45 expression) and assess our transduction efficiency (eGFP expression).     The first ...detected through a cell surface label. The viral vector also encodes for eGFP, which serves as a transduction...rest of our panel. Lastly, we want to detect the expression of our protein of interest: CD45. I am using ...

...2015). The dual vector system uses S. pyogenes Cas9 (SpCas9), using one vector to express SpCas9, and another...delivering Cre and sgRNA to a Cre-dependent mouse or sgRNA to a constitutively Cas9-expressing mouse. Described...luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone with luciferase-2A-Cre recombinase Vectors for brain...mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer modeling Using Cas9 mice, Platt...pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR This plasmid contains two expression cassettes, Renilla luciferase-2A-Cre recombinase...target site sequence (20bp). CRISPR-Cas9 Cre expression vectors for genome editing in the brain Using Cas9...

...Collections Gene Vector Core Viral Vectors Gene Vector Core Viral Vectors Lentiviral Transfer Vectors Adenoviral... transfer vectors and generating new viral vectors. Download the Lentivirus Transfer Vector Production...Lentiviral Vector ID Purpose pCDH-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter when Cre is expressed...by another vector pCDH-EF1-DIO-tdTomato 72254 Expresses tdTomato under EF-1 promoter when Cre is expressed...promoter when FLP is expressed by another vector pCDH-EF1-Fon-tdTomato 72261 Expresses tdTomato from the ... promoter when FLP is expressed by another vector pCDH-EF1-FLPe 72262 Expresses FLPe from the EF-1 promoter...lentiviral and adenoviral vectors read our Lentiviral Vector Guide and Adenoviral Vector Guide Find more plasmids...

...2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR/Cas9 Toxicity in Human...genome editing, activating gene expression, repressing gene expression, and much more. But there’s one... means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems prevents... off-target effects Prolonged Cas activity can increase the chance of off-target editing particularly ...

...Bradbury et al., 2018), and hybridomas can lose gene expression of the mAb-encoding genes, die, or fail to revive...research materials, such as plasmids and viral vectors. However, it typically takes a few years for a ...antibodies are not monospecific: Hybridomas frequently express additional functional variable regions. mAbs 10...these issues by creating immortal antibody-producing cell lines (hybridomas) that secrete antibodies in ...Neurotechnologies® (BRAIN) Initiative, will allow Addgene to create a much needed open-access recombinant antibody/...School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...reagent batch. To supplement this, Addgene will be creating a crowdsourced Data Hub (built off of the concept...

...genes is from Proteobacteria to mammalian expression vectors. And more broadly, their data revealed that...a distantly related organism, the gene may not express well because of these different codon usage biases...closely resembles that of the host organism, it may express protein more efficiently. To classify gene sequences...rare codons, sometimes intentionally modulating expression in ways we don’t quite understand yet,” Kunjapur...synthetic. They also found out that the most common expression system within the Addgene repository is mammalian...series of simulations and machine learning, they created a classifier that predicts whether a gene was natural...

...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different promoters...reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications...the Wu Lab is working to create a three-dimensional atlas of protein expression in the adult mouse brain...activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive element...enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can increase on-target .... The Ting lab recently made two deposits that increase the utility of this technique.  By engineering... O’Neill The Cohen lab and colleagues recently created a system for recording the transcriptional history...

...FLEX-tdTomato-T2A-SypEGFP-WPRE Syn-driven, Cre-dependent Cre-dependent expression of cytoplasmic tdTomato and synaptophysin-EGFP...2A-Synaptophysin-mRuby Syn-driven, Cre-dependent Cre-dependent expression of membrane-bound GFP and synaptophysin-mRuby...124364 pAAV-FLEX-DTR-GFP CBA-driven, Cre-dependent Cre-dependent expression of diphtheria toxin receptor fused...pAAV-FLEx-ER-TurboID CMV-driven, Cre-dependent Cre-dependent expression of ER-localized TurboID (with HA...pAAV-FLEX-P301L Tau EF1a-driven, Cre-dependent Cre-dependent expression of human 2N4R microtubule-associated...pAAV-DIO-BACE-HA Syn-driven, Cre-dependent Beta-Secretase to be expressed (1:1) with either ATLAS protein... Viral Vector Packaging Service AAV Molecular Tools Viral Vector Packaging Service: Molecular...

...Color-flipping switch. Expresses NLS-mCherry in the absence of Cre, and expresses NLS-EGFP in Cre-positive cells...AAV-pgk-Cre PGK Cre expression Recombinases Patrick Aebischer 55632 pAAV-Ef1a-mCherry-IRES-Cre EF1a Cre...physically-separate mCherry expression Recombinases Karl Deisseroth 55636 pAAV-EF1a-Cre EF1a Cre expression Recombinases...-tagged Cre expression Recombinases James M. Wilson 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Syn Cre expression...pENN.AAV.CamKII 0.4.Cre.SV40 CamKII Cre expression Recombinases James M. Wilson 107738 pAAV-hSyn-Cre-P2A-dTomato...pAAV-hSyn-Cre-P2A-dTomato Syn Cre expression with simultaneous dTomato expression Recombinases Rylan Larsen 107788 AAV.rTH.PI.Cre.SV40...EF1a Flpo expression Recombinases Karl Deisseroth 55638 pAAV-EF1a-vCre EF1a VCre expression Recombinases...

...can be expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate vector. ...CRISPRainbow gRNA will permit labeling, but a newly expressed standard length gRNA would “switch” the system...scaffold to enhance complex assembly, thereby decreasing the amount of background fluorescence from unbound...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...was fused to a different fluorescent protein to create three distinct colors. The specificity of these...distinct hairpin sequences produces white light. Created with BioRender.com. With this rainbow of colors...combination, for example, with a far-red FP, would increase the available colors to 15! They also envision...

...interest. In our studies, we used AAV viral vectors to express hM4Di in neurons of rhesus monkeys or mice...successful expression of the protein of interest, an alternative could be bicistronic expression of the fluorescent...the fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other...proteins provide a versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore...can be a roadblock, rather than an aid, in the expression of the protein of interest. The green fluorescent...’ tags to identify proteins Given the limited expression of hM4Di‐mCherry at the plasma membrane in monkey... mCherry with the HA tag resulted in abundant expression of hM4Di in plasma membranes in both species ...

...instance, if you were cloning a gene into an expression vector, you would want the start of the gene to end...Your Insert and Vector by Gel Purification     Now that you’ve cut your insert and vector, unfortunately...unsuccessful 1), in a vector alone plate with many more colonies than the vector + insert plate (unsuccessful... ligation step. 3. Ligate Your Insert into Your Vector In the ligation step, you mix your purified, cut...Resources on Addgene.org Find the Perfect Empty Vector for Your Cloning Experiments Browse Our Molecular... 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert...

... compared to the E. coli strain expressing the empty vector. And thus was born ELSA - our happy lil’ cold-tolerant...decided to focus on a special class of enzymes expressed by psychrophilic (cold-loving) bacteria. These...recombinant psychrophilic enzymes for the host The expression of recombinant proteins for industrial or research...mesophilic, for example E. coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are...enzymes face very similar issues when forced to express in mesophilic host systems. The so-called “regular... to grow at low temperatures suitable for the expression of psychrophilic proteins. The primary problem...Factor), that would allow the stable folding and expression of recombinant psychrophilic proteins. As the...

...years, he and his colleagues built a Molecular Expressions collection including some 3,300 plasmids along... Baird said. “They are all designed in the same vector backbone with very similar restriction enzymes....shutter its doors. “We wanted to give all of these vectors to the scientific community and contribute in a...proposition, explained Michelle Baird, who has created plasmids as a technician in the Davidson lab for...prompted him to deposit the tools he and his team created with the research community at large, "[Fluorescent...

...85040 pK170.AAV-TRE-Cre-WPRE (Supernova) AAV vector for dox-inducible expression of Cre recombinase. Takuji...63704 pRetroX GFP T2A Cre Retroviral vector for dox-inducible expression of GFP T2A Cre recombinase. Floris...Neo-TRE-CMV-Cre-rtTA Dox-inducible expression of Cre recombinase. Madeline Lancaster 198752 DiLiCre 2.0 Dox-inducible...popular empty backbones, expression vectors for transactivators, and vectors for other applications, or...Lentiviral Tet-Off vector for repressible expression with blasticidin selection. *Note: this vector contains an...PB-EF1a-TetOn3G PiggyBac vector expressing Tet-On 3G transactivator constitutively expressed by EF1a promoter ...Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Tetracycline Off Tetracycline...

...individually expressed proteins run in separate lanes. B: individually expressed and coexpressed proteins ...through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either 10, 30, 50...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure... Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when encoding multiple guides. While...opportunity for lab members or students new to protein expression and purification. Figure 3: The Penn State...Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60, 80 and 100 kD proteins...expanded the genetic screening toolkit by optimizing a Cas12a-based method for screens. Cas12a’s ability ...

...excellent start (#22988, #137884). Both vectors allow the creation of amiRNAs based upon miRNA precursors...transgene. Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region...considerations, such as the starting material (DNA-based vector or purified RNA) and the initial state of the dsRNA... easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This method is facilitated...construct has unique features, such as inducible expression of the hpRNA and/or fusing your gene target sequence... Although downstream work is required, several vectors can be found at Addgene that could serve as an ...ones? Here, I’ll cover some screening methods to identify them. Visual screening Regardless of the RNAi-trigger...

...then easy to package in low-capacity vectors like AAV, a popular vector due to its low immunogenicity. But...in size and are easy to package in low-capacity vectors! These small, but mighty type VI-D enzymes are ...collateral cleavage does not occur when Cas13d is expressed in a mammalian system. Since Cas13d is functionally...RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased EsCas13d...Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR repeat array near a putative... WYL-domain-containing accessory protein WYL1 increases RspCas13d and EsCas13d cleavage activity The majority...

...in a single vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from...III transcription, potentially increasing the levels of gRNA expression. Since tRNAs are among the most... confirming gRNA cleavage, they examined gRNA expression levels and indel generation for two PTG constructs...protoplasts. Compared to matching traditional gRNA-expressing constructs, transcript levels of PTG1 and PTG2...Xie et. al are specifically designed for plant expression, PTGs will likely be adaptable to many different...capacity. Still confused about the various tRNA expression strategies? No worries, I've summarized the three...research? Check out the PTG plasmids or plant expression systems available from Addgene. If you have any...

...Model Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid System ...Phillips made the decision to release the K. phaffii expression system to the general research community in the...different uses for the phaffii-lous strain. Protein expression   K. phaffii first gained attention for its powerful...This gene promoter, AOX1, was co-opted for the expression of recombinant proteins by the pharmaceutical...pharmaceutical and biotech industries as a protein expression workhorse. This yeast can also give more protein bang...Saccharomyces cerevisiae to human: The important gene co‑expression modules. Biomedical Reports 7.2 (2017): 153-158...protein secretion, making harvesting recombinant proteins a cinch. This is because it doesn’t secrete its ...

...Cre-expression Cre Wilson 107738 pAAV-hSyn-Cre-P2A-dTomato Syn Cre-expression Cre Larsen 140135 pAAV-EF1a-iCreV...pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 Syn Cre-EGFP expression Cre Wilson 105550 pAAV.GFAP.Cre.WPRE.hGH GFAP Cre-expression... Viral Vector Packaging Service AAV Caltech Systemic Capsids Viral Vector Packaging Service... a new window) . Viral Vectors 101: Systemic Capsids PHP.eB These viral vector preparations were produced...Activator, Cre-dependent DREADD Roth 44362 pAAV-hSyn-DIO-hM4D(Gi)-mCherry Syn Inhibitor, Cre-dependent ...sensor, Cre-dependent GCaMP Kim , GENIE 104492 pGP-AAV-syn-FLEX-jGCaMP7f-WPRE Syn Calcium sensor, Cre-dependent...sensor, Cre-dependent GCaMP Kim , GENIE 104496 pGP-AAV-CAG-FLEX-jGCaMP7f-WPRE CAG Calcium sensor, Cre-dependent...

...interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine with N-terminal...Cyan Mammalian Expresses mammalian optimized Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest...Cyan Mammalian Expresses mammalian optimized CyPet pCyPet-His Cyan Bacterial Expresses CyPet with C-terminal... Mammalian Expresses mammalian optimized YPet pYPet-His Yellow Bacterial Expresses YPet with C-terminal...Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover Green Mammalian Expresses Clover...Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between ECFP and ...monomeric Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...

...of robust expression of high G+C content genes, as validated through monitoring the expression of fluorescent...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check... be seen through the success of the change in expression of GFP, for the stop codon should have changed...different biochemical secondary energy sources to increase protein yields from 36 μg/mL to 266 μg/mL. The...oxytetracycline enzymes. Next, this system was used to recreate natural biosynthetic pathways. Biosynthetic pathways...

...viral vectors.  Caspedia: A data repository for Cas enzymes.  Open repository of CRISPR screens Cellosaurus...enzyme cut sites Open reading frame finder  Addgene vector database: Find sequences for plasmids assembled... sequence and functional information, such as expression patterns, post-translational modifications and...plasmid analysis tool including editing, annotating, creating maps, and more. This tool is maintained by M. ...University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA. ...

... to cut targeted genes, creating mutations that prevent their expression. The knockout cells can be used...Library, whose acronym means Modular, NGS-trACKable ExpRession Element. This library consists of broad host ...interest are fused to anchoring proteins to be expressed on the surface of a host cell or virus, such as...Additional resources on the Addgene blog Viral Vectors 101: Preparing Pooled Libraries Pooled Library ...genome at once! Knockout screening with CRISPR libraries In CRISPR knockout screening, sgRNAs guide Cas9 to...: Schematic of CRISPR knockout screening with a pooled library. Created with BioRender.com. The popular...libraries called Julianna, for mouse screening, and Jacquere, for human screening. These new libraries have improved...

...gene expression. The regulatory element is cloned upstream of the luciferase gene, and expression of luciferase...backbones with the luciferase gene, a variety of expression vectors, and literally thousands of reporter constructs...wavelength of light they emit or to optimize their expression in mammalian systems. Our luciferase collection...luciferase assays have become a gold standard in gene expression analysis to investigate the effect of regulatory...interactions Split luciferases, such as NanoBiT, are expressed in two subunits and only catalyze a luminescent...reaction catalyzed by luciferase, releasing light. Created with BioRender.com. Bioluminescent reporters...Peak emission (nm) Size (kDa) Substrate Cofactors Secreted North American firefly (Photinus pyralis) Natural...

...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...Non-autonomous transposons, however, need another TE to express a transposase or reverse transcriptase so they ...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...

...cells express iCre with a mitochondrial anchor and a TEV cleavage site. Recipient cells express DsRed ...Once the recipient cells take up the Cre-tagged mitochondria, Cre is released, translocates to the nuclease...killswitch (GFP-nb–KS) is delivered using a vector that also expresses mCherry to mark transfected cells. The...potent effect, increasing oxygen consumption and upregulating other genes normally expressed in BAT. When...cell-surface proteomes (Wu et al., 2025). The team expressed extracellularly-anchored horseradish peroxidase...mitochondria (Hoover et al., 2025). MitoTRACER relies on a Cre-lox switch to turn recipient cells from red to green...nuclease, and excises the DsRed-STOP to trigger expression of EGFP. Figure 2: Schematic of the MitoTRACER...

...63704 pRetroX GFP T2A Cre CMV/MSV Tetracycline inducible expression of GFP T2A Cre fusion in mammalian ...GFP MoMLV U6-driven plasmid for shRNA expression with GFP expression. See plasmid 8452 for puromycin resistance...Luciferase MSCV Plasmid for transgene expression with luciferase expression. Scott Lowe 9044 pMIG MSCV Plasmid...Plasmid for transgene expression with GFP expression. See plasmid 12282 for the addition of WRE, a post-transcriptional...27995 TtRMPVIR CMV/MSV Tet-regulated expression of shRNA. Expresses rtTA. See Zuber et al. (2011) for additional...pLncEXP CMV/MSV lncRNA expression plasmid. Lei Sun 60683 pLXIN-Luc MoMSV Stable expression of luciferase in ...pMD2.G VSV-G-expressing envelope plasmid. Didier Trono 35617 pCAG-Eco Ecotropic MLV expressing envelope plasmid...

...LinkedIn.   References 1. Yin, R., et al. "Gene expression profiling analysis of bisphenol A-induced perturbation...cytoskeletal disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied...with the Agarose Method Plasmids 101: Mammalian Vectors Resources on Addgene.org Find mammalian plasmids...plasmids for your research Find viral vectors for infecting mammalian cells ...making their surrounding media acidic through the secretion of lactic acid (a by-product of cellular metabolism...

...plant We developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome...genome. We constructed three versions of the vector (pKABE6.3, pKABE7.9, and pKABE7.10) so that the user...are better suited (Gaudelli et al., 2017). Our vectors have two Bsa1 sites to clone the single guide RNA...scaffold) in a pol-III (OsU3) promoter driven expression system. We use a single polycistronic tRNA-gRNA... genomic site simultaneously.     These binary vectors could be used to generate stable genome edited ...Agrobacterium-mediated transformation. We also constructed smaller vectors suitable for transient assays using protoplast ...cytosine deaminase with Cas9 nickase (nCas9). nCas9 creates a nick in the DNA by cutting only a single strand...

...determined by comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry. ACE was...mCherry and a T2A site and expressed in cells. mCherry is constitutively expressed to identify successfully...that can be repaired by NHEJ restoring mCherry expression. Cells can then be sorted using Fluorescence-...BFP variant, a BE4am-Cas9 base editor, and sgRNA vectors for the BFP-to-GFP conversion and to target specific...integrated it into human and mouse cells that expressed either Cas9 or an optimized cytidine base editor...antibiotic resistance markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing... of a mutated start codon to initiate protein expression, it can be used for an array of detectable proteins...

...other molecules to host organisms. For instance, vectors derived from the Agrobacterium Ti virulence plasmid...especially when combined with natural inducible expression systems that allow tuning of metabolic pathways...developing new cloning techniques based on these vectors (Shareck et al., 2004). Plasmid categorization..., Y., Lee, B., & Miguez, C. B. (2004). Cloning Vectors Based on Cryptic Plasmids Isolated from Lactic ...Figure 1: Common functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids...resistance genes to help cells survive their own secretions. Pro tip! Historically, colicin-producing plasmids...years, and new classifications are still being created as whole-genome sequencing becomes more relevant...

...plasmids with low to medium copy numbers can still express massive amounts of protein given the proper promoter...the table below for more details.   Common Vectors Copy Number+ ori Incompatibility Group Control ...Relaxed This table defines common cloning vectors, their copy number, ori, and incompatibility groups...R6K, and 15A. Not all origins of replication are created equal. Some will produce many plasmid copies and...because they will compete for the same machinery, creating an unstable and unpredictable environment. As ...plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain Most E....

...use plasmids from Addgene to construct NFIX expression vectors as well as reporters of NFIX function. They...interactions and how these interactions regulation gene expression. Recent studies have shown that the conditions...delete the functional portion of the Nfix gene using Cre recombinase and to generate Nfix-negative neural ...interactions and how these interactions regulation gene expression.     ...burden of immune symptoms on CDG patients and to increase their quality of life by reducing their risks ...ligand binding sites in T4 lysozyme by deficiency-creating substitutions." Journal of molecular biology 277.2...

...My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, ...shell, or capsid; 2) the pHelper plasmid which expresses adenovirus genes which help mediate AAV viral ...Production of recombinant adeno-associated viral vectors for in vitro and in vivo use. Current protocols... out what makes AAV a versatile tool for gene expression in mammals Use the PEI calculator for planning...transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with a SmaI or XmaI restriction...duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary. To save time...mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial pellet...

...in patched cells expressing wild type CoChR, these APs are absent in soCoChR-expressing cells. Finally,...C-terminus of the protein. Unlike the diffuse neuronal expression observed with wild type CoChR, soCoChR localizes...stimulation are largely absent in the soCoChR-expressing system. Taken together, Shemesh et. al. describe...Pseudomonas putida CBB5 and has been tailored for expression in E. coli. This operon is under the transcriptional...strong constitutive promoter (BBa_J23100) and expresses N-demethylases and other required enzymes, including...Moreover, a caffeine “addicted” E. coli strain expressing this operon can serve as a living biosensor for...FRET reference page for many other common FRET vectors. Murakoshi H, et al. Sci Rep. 2015. Pubmed PMID...

... Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into...confirm the presence of an insert in a particular vector by excising the insert from the backbone. To do...will need to know both the approximate size of the vector backbone as well as the predicted size of the insert...time-intensive, so we recommend a variety of ways to screen and verify your plasmids. Here, we'll cover restriction...500bp to 10kb, with the 3.0kb fragment having increased intensity to serve as a reference band. The uncut...

...Fluorescent Protein (GFP) Fluorescent Protein Guide Expression Vectors from the Berkeley QB3 MacroLab Eric thanks...box of tags and fusion proteins to aid in the expression and purification of recombinant proteins. Tags...purification to get your protein as pure as possible? Vectors that offer different combinations of tags are readily...Biochemists and molecular biologists who need to overexpress and purify proteins can face any number of technical... some of the hurdles to overcome in order to overexpress a recombinant protein? It is not generally in...in a cell’s best interest to overexpress a protein. Energy and cellular resources are being spent to make...stabilization, as can maltose-binding protein (MBP). Overexpression can also lead to insolubility, and aggregated...

...epitope tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster...antibody-based detection, smFPs create much brighter signals for weakly expressed proteins than standard tags...cytotoxicity, low immunogenicity, and the long-term expression of AAV delivered genes. AAV serotypes have been.... Thanks to this work it is now much easier to express molecules in your favorite neuronal population ...insufficient for detection when the POI is weakly expressed. As these epitopes tags have no intrinsic visual... be fused to scaffold proteins in order to be expressed stably in neurons. As compared to epitopes tags...labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult to track ...

...protein of interest into a vector encoding a fluorescent tag. Once expressed as a fluorescent protein fusion...bacterial components. For example, the Rainbow Vectors from the Mariette Barbier Lab can be used to fluorescently...Barbier, Mariette, and F. Heath Damron. "Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling... to its final destination. As an example, most secreted proteins are produced in the endoplasmid reticulum...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ... they are finally degraded (2, 3). Once you've created your fusion protein, it's important to validate...subcellular structure or protein complex. By coexpressing both the marker protein and the protein of interest...

...compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors - Epitope ...lentiviral vector for ORF expression pDest-565 - Gateway destination vector for bacterial expression; His,GST...Empty gRNA Expression Vectors page Cre-lox Site-specific recombination See our dedicated Cre-lox Plasmids...gene expression vector URA3 Yeast pAG306GAL-ccdB - Yeast Gateway expression vector (see...Inducible Expression page Bacteria Lac, T7, araBAD, trp mTagBFP2-pBAD - Protein expression vector with C-...versions of pFastBac LIC vectors and pFastBac Dual LIC vectors for insect expression Fluorescent proteins ...Bacterial expression of EBFP2 blue fluorescent protein 35S-eGFP-nosT - Transient expression vector for eGFP...

... library via a lentiviral vector to cell lines that either stably expressed Cas9 or that had been lentiviral...control for many of the effects of gRNA and Cas9 expression, they fail to account for the effects of Cas9...sites should control for the effects of guide expression and dsDNA breaks. Figure 2: Non-targeting...biological process. These screens provide an alternative to genome-wide RNAi screens, which although highly...similar pitfalls exist for CRISPR screens and how best to design these screens to controls for flaws. Recently...CRISPR knockout screen to analyze the following unanswered questions about CRISPR screen design. Unanswered...CRISPR/Cas9 screen design Are non-targeting guides an appropriate control? Many CRISPR screens use non-targeting...

...fluorescent proteins, reporters, biosensors, and Cre expression plasmids. Large collections or kits are described.... pGCS Vector Kit - Hai-Ning Du Lab. A set of pCS2+ backbone-based Gateway destination vectors (pGCS),...which an exogenous mRNA can be injected and expressed. EXPRESs Plasmids for Zebrafish Receptors and Zebrafish...in zebrafish. Zebrafish expression atlas - A zebrafish-specific gene expression database that provides ...resistant cell types. CreLite - George Eisenhoffer Lab. An optogenetically controlled Cre/loxP system that ...A useful “starter kit” for screening fluorescent protein fusion expression in organisms or cells in which...toolbox of genetic methods, such as Tol2 transposons, Cre-mediated site-specific recombination, morpholinos...

...CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain GABAergic interneurons...combination with Cre-dependent transgenic mouse lines or co-infection with Cre dependent viral vectors. Jonathan...pAAV-AiE0452h-minBG-iCre(R297T)-BGHpA (Alias: CN3781) Enhancer AAV for Cre recombinase expression in striatal ..._3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN3779) Enhancer AAV for Cre recombinase expression in striatal ...pAAV-AiE0450h-minBG-iCre(R297T)-BGHpA (Alias: CN3778) Enhancer AAV for Cre recombinase expression in striatal ...version of DLX enhancer). AAV vector for strong & rapid SYFP2 expression in forebrain GABAergic interneurons...pAAV-AiE0441h_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN4825) Expression of iCre(R297T) in striatal medium...

... channels, such as ChR2 and NpHR, can even be expressed simultaneously and still be individually activated... like spontaneous swimming. To do so, the lab expressed the LiGluR opsin in different subsets of spinal...optogenetics blog posts Read blog posts about viral vectors Subscribe to the Addgene blog Resources on Addgene.org...spinal cord neurons using the Gal4/UAS system. This created fish lines with different groups of neurons that...

... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core. Transfections...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Transducibility Some viral vectors are tested in vitro and in vivo for gene expression and/or function. These....1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct fluorescence...Empty vector particles can be identified after negative staining and appear darker than full vector particles...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...

...to induce the expression of a Cre recombinase contained within the knock-in. Including Cre-mediated marker...kilobases of additional sequence being introduced. Cre-mediated excision of the selective cassette minimizes...experimental manipulation. In some cases, injection of Cre is required, however the development of the self-...modular toolkits build single plasmid targeting vectors that encode the gRNA transcript and the repair ...using modified Cas9 proteins to regulate gene expression (CRISPRi/CRISPRa), visualize specific genomic....114.170423 Schwartz ML, Jorgensen EM (2018) SapTrap vectors for introducing point mutations with unc-119+ selection...variable amounts of experimental manipulation and screening.  A co-selection strategy for CRISPR editing in...