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We narrowed to 370 results for: Cre expression vector

Showing: 161 - 200 of 370 results

  1. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

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    Blog Post
    ...transdifferentiated in vivo to pancreatic β-like-cells with the expression of pancreatic and duodenal homoeobox.... Ferber, S., et al., Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver...four reprogramming factors can be delivered and expressed in multiple somatic cells via various methods....transdifferentiation is usually carried out by expressing transcription factors specific to the lineage...and in vitro methods are similar except that the vectors carrying the transdifferentiation factors are directly... (iPSCs) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A, 2011. 108(34): p. 14234...., Human induced pluripotent stem cells free of vector and transgene sequences. Science, 2009. 324(5928...

  2. Mouse Modeling, Part 2: Breeding and Crossing Mice

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    ... that expresses Cre with a strain that expresses a floxed gene of interest. Cre may be expressed globally...heterozygous for each mutated allele.  Cre-lox expressing strains Cre-lox strains are a common example of... in mice expressing both Cre and LoxP occurs. But how are these mice made to begin with? Cre-lox mice ...tissue-specific manner (with expression controlled by a tissue-specific promoter). And, Cre may also be either ...constitutively expressed, or inducible (this is explained more in Part 1 of this series). In any case, Cre is a ...which you’ll want to genotype your mice for expression of Cre, loxP, and your gene of interest. To ensure...site, your control should be littermates that express Cre but not loxP. With this two-part guide to mouse...

  3. Hot Plasmids: Winter 2025

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    ...cell-derived neurons expressing ASAP5-Kv. C) Cultured rat hippocampal neuron expressing pan-membrane ASAP5...Each plasmid expresses all the components of the biosensor in optimal proportions from a vector backbone ...Figure 2), the authors developed a cost-effective expression and labeling approach using these constructs ...protein of interest can be inserted. B) Protein expression and labeling procedure. Adapted from Smith et..., and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only for future... The authors have deposited AAV plasmids for Cre-dependent and -independent versions that are trafficked...recently developed a collection of BRET-based ONE vector G-protein Optical (ONE-GO) biosensors (Janicot ...

  4. Modulate the Activity of 17 Signaling Pathways with One Kit!

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    ... cDNAs into a single consensus lentiviral expression vector and attached a DNA barcode to each to make...cDNAs) representing core nodes that, when over-expressed, constitutively activated or inactivated the pathway...other researchers, and cloned them into lentiviral vectors to generate a kit which is now available at Addgene...make the library usable for pooled screens. Applications of the cancer pathways ORFs kit Wood and his colleagues...colleagues developed the kit for use in screening for pathways that drive resistance to drugs in vitro...the only way to use the kit. “You could use it to screen for pathways that confer any phenotype of interest...drug resistance,” he said. “It could [be used to screen for] morphological features, migration, or survival...

  5. Working with Nuclear Receptors

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    ... a vast number expression and reporter plasmids and the free exchange of these vectors between laboratories... ligands remain unknown. Together, NRs govern expression of genes involved in a broad range of reproductive...efficient and timely activation or repression of gene expression (see www.nursa.org for a curated list of coregulators...ligand, cell type, and promoter, organizing the expression of functional groups of genes involved in the...Our Q-PCR datasets, for example, document the expression patterns of nuclear receptors across multiple...researchers with a one-stop shop for plasmids and vectors, and making them available to all researchers for..., B. W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators. Cell 108...

  6. Another Pathway into Cells: iTOP

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    ...in which transient protein expression has a long-lasting effect, such as Cre- or Cas9-mediated recombination... mutations and gene expression changes elsewhere in the genome. Second, expression is by nature transient...control could enter cultured cells and activate expression of a luciferase reporter. CPP was not required...tested iTOP in a series of primary cell types using Cre and various loxP reporters. In both mouse and human...stem cells transduced with iTOP resulting in GFP expression. Image courtesy of Diego D'Astolfo. CRISPR...potential application of iTOP Having tested iTOP using Cre-lox recombination, D’Astolfo et al. were eager to...causes a cell to take up, and continued protein expression would require multiple transduction events. For...

  7. Plasmids 101: Inducible Promoters

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    ...inducible system is found in many bacterial expression vectors. Negative inducible promoter pBad is another...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...Temperature inducible promoters Temperature sensitive expression systems are typically less leaky than chemically...chemically induced promoters; they show near-zero expression at regular temperatures but can be induced by ... promoters, in which a gene of choice is only expressed following exposure to a brief heat shock. In the...Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering in species like...promoters Light is another way to activate gene expression, and two-component systems used in synthetic ...

  8. Plasmid Tools for Microbiome Studies

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    ...can also detect expression of toxin mRNA from Clostridium difficile. As mRNA expression cannot be identified...2009). The vector retains the oriT for delivery into recipients by conjugation. The labs screened for the...upon the Yeast Toolkit by replacing the backbone vector’s high-copy number bacterial ColE1 origin with the...resistance markers, promoters, and inducible gene expression systems in bee gut microbiome species such as... or a bacterial microcompartment protein. The expression of this fusion is under control of the arabinose-inducible...Cells are first induced with arabinose to begin expressing the SAP-RFP fusion. Then, the cells are washed...of cells containing the particle exponentially decreases over time and the number of elapsed generations...

  9. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    ...Addgene.org Browse Our Viral Vector Plasmids Read Our Lentiviral Vector Guide Read Our AAV Guide  ...producing infectious virus (similar to lentiviral vectors) because they do not encode all of the necessary...plasmids allows researchers to partially control expression levels of each gene by altering the amount transfected...RNA polymerase gene or using cells that stably express T7 (e.g. BSR T7/5 cells) (1). We hope that minigenomes...on the Addgene Blog Read Other Posts on Viral Vectors Check out Our Scientist's Guide to Ebola Contribute... viral vector...This post was contributed by guest blogger Tessa Cressey. The highly pathogenic Ebola virus belongs to the...

  10. Overcoming the Challenges of Lentiviral Production

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    ... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges... a usable titer, or fine-tuning selection and expression in your target cell line, researchers often find...transcriptional activation system to drive high levels of expression of certain viral components; according the manufacturer...determination of the packaging limit of lentiviral vectors. Hum Gene Ther. 2001 Oct 10;12(15):1893-905. PubMed...Tips for titering your lentivirus Visit the viral vector featured topic page Resources on Addgene.org ...Guide Find lentivirus plasmids Browse all viral vectors ...consider scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size. Opting ...

  11. Genome-wide Screening Using CRISPR

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    ...lentiviral transfer vectors to create the CRISPR library. CRISPR libraries have been created to knock out, ...deliver a pooled population of gRNAs to Cas9 expressing cells. Pooled lentiviral CRISPR libraries (referred...population of gRNA-containing lentiviral transfer vectors, each targeting a specific gene within the genome...) to be used to generate lentivirus (B). Cas9-expressing cells or wild-type cells are treated with lentivirus...genome-wide screens? The goal of a genome-wide screening experiment is to generate and screen a population...the steps involved in a CRISPR screen? Performing a forward genetic screen using CRISPR libraries is a multi-step...sorting and screened for a particular phenotype. For example, mutant cells can be used in drug screens to identify...

  12. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

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    ...cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation sequencing...the stop cassette is removed, the effector is expressed. By truncating one of the recombination sites ...calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin is required for virulence...4X CDRE repeat. In addition to showing that this vector is suitable for flow cytometry and fluorescence... single molecules in real-time. Find plasmids expressing tTALE-FP Shin et al., Sci. Rep. 2019. https:/... Figure 1: Initial steps in the CRISPR-pooled screening using sgRNAiBAR. Image from Zhu et al., 2019.  ... Peking University has developed a new CRISPR screening method that makes use of guide RNAs with internal...

  13. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    ...categories Expression: Select the model system for your research (e.g. Mammalian) Vector Type: What type...Addgene is currently distributing 94 plasmids expressing validated antibodies for neuroscience research...example I wanted to find a lentiviral plasmid to express ubiquitin under control of a CMV promoter. Starting...but I can’t browse through 300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates...Identify a specific marker protein Use: Similar to “Vector Type” you can filter by experimental purpose (e.g...more about this!). I’ve assembled a series of screenshots to highlight changes to both the design and functionality...NeuroMab Collection for details). The following screenshots show the range of different terms we can use ...

  14. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

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    ...the Krogan lab at UCSF generated lentiviral vectors expressing SARS-CoV-2 open reading frames and identified...plasmids expressed 26 of the 29 SARS-CoV-2 proteins which all but one are cloned into pLVX vectors (for lentiviral...provide synthesized SARS-CoV-2 expression plasmids available for many expression systems: bacterial, yeast,...C-terminal deletion in the spike protein to improve expression and pseudotyping efficiency. This plasmid was...enzymes. Plasmids in this toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase...-CoV infection and deposited plasmids for the expression of cathepsin S, L, and B. This became a great...human cells. Addgene’s mammalian target plasmids express these and other targets such as TMPRSS2, FURIN,...

  15. Overcoming the AAV Size Limitation for CRISPR Delivery

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    ...Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of...activation. Split inteins have also been used to express base editors. References Chew WL, Tabebordbar...HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038...genome engineering: An adeno-associated viral (AAV) vector toolbox. Biotechnology Journal 9:1402–1412 . https...Editing: Strategies and In Vivo Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell...

  16. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

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    ...Practical Guide: Using pK-TnpB1 and pK-TnpB2 Vectors The vectors pK-TnpB1 and pK-TnpB2 are designed for plant...the digested vector. Step 4: Ligation Ligate the annealed oligos into the digested vector using your ...efficient delivery into cells, particularly via viral vectors. While viral delivery is a highly effective method...genome editors to date. We have developed multiple vector versions for TnpB-mediated genome editing, and ...delivery of editing reagents into cells. Viral vectors, because of their natural ability to infect cells...approval and public acceptance. Recently, viral vectors have been successfully used to generate gene-edited...al., 2025). However, a key limitation of viral vectors is their restricted cargo capacity, which makes...

  17. Same Addgene, New Look - Why We Redesigned Our Homepage & Mascot

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    ...’ the homepage is still important. It needs to express value, clearly communicate exactly what a site ...wanted users to be able to find plasmids and viral vector tools more easily. In the redesign, the new search...One big feature upgrade? Blugene now has a full expression. You’ll mostly see a friendly smile, but occasionally...you come to Addgene to find the plasmid or viral vector for your next experiment, that you can find what...Addgene.  Addgene's new homepage While users are increasingly accessing websites other than through the ‘front...

  18. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    ...same strain. MazF expression is under control of the AOX1 promoter whose expression is induced by methanol...that are used to drive high protein expression in vivo. Expression plasmids are often lost without plasmid...Kaper used the hok/sok system to create strains that can stably express antigens in sufficient quantities...as a counterselectable marker in the destination vector. Successful cloning occurs when the ccdB gene is...methanol. Once linearized, the delivery vector can be integrated into the genome to introduce the genetic...plasmid R1: Sok antisense RNA regulates hok gene expression indirectly through the overlapping mok gene."...James B. Kaper. "Use of stabilized luciferase-expressing plasmids to examine in vivo-induced promoters...

  19. Pushing the Limits of DNA Assembly

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    ...homology. Producing yeast expression cassettes Traditionally, generating expression cassettes for yeast transformation... mutations Cloning sgRNAs Production of Yeast Expression Cassettes Generating constructs with multiple...PCR to generate the mutation and linearizing your vector (preferably by PCR) for DNA assembly. Further, ...simple as mixing fragments together with linearized vector for seamless joining of multiple mutations spread...physically joining multiple fragments of DNA to create a synthetically designed DNA sequence. There are...fragment joining, but this is fine for its goal of creating a choice of standardized constructs and tools ... regions. Golden Gate Assembly also allows the creation of standardized DNA constructs, but its use of...

  20. OCaMP: A New Calcium Indicator for Neural Imaging

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    ...fluorescence responses to increasing numbers of evoked APs in neurons expressing OCaMP or jRGECO1a. Reproduced...electrophysiological recordings (lower trace) using OCaMP expressed in mouse cortex. Bottom: Zoomed-in view of traces...FLP)-OCaMP-WPRE pAAV Ef1a pAAV-hSyn-FLEX(cre)-OCaMP-WPRE pAAV hSyn pAAV-CMV-OCaMP-WPRE...Additional Resources on the Addgene Blog Viral Vectors 101: Calcium Sensors New Neuroscience Tool: The...Fluorescent Proteins: Biosensor Plasmids In-Stock AAV Vectors: Biosensors AAV Packaged on Request  .... These industrial fiber lasers are becoming increasingly common due to their low cost and high power,...

  21. What's New in CRISPR - March 2020

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    ...fluorescently labeled piggyBac vectors to deliver uniform and sustained expression of multiplexed gRNAs in human...CRISPRa depended on continued expression of dCas9 and gRNAs whose expression varies depending on transgene...The lab expressed both dCas9 and the ctrA sgRNA and saw that the levels of ctrA mRNA decreased after 20...demethylation Optogenetic repressors of gene expression in yeast Megan McClean’s lab generated an optogenetic...optogenetic toolkit that controls repression of gene expression in Saccharomyces cerevisiae. The system consists...the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the NLS is inaccessible and the... in Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo et al., 2020...

  22. Viral Vectors 101: Voltage Indicators

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    ...specific cell types is where viral vectors become invaluable. Viral vectors are mechanisms by which genetic...Resources on the Addgene blog Viral Vectors 101: Chemogenetics Viral Vectors 101: Optogenetics Fluorescent...Sensors This system consists of a genetically expressed enzyme, typically on the cell surface, which can...fluorescence or voltage indicators.   Viral Vectors and Voltage Indicators As previously mentioned,...hydrolyzes a precursor dye to make it more lipophilic, increasing its affinity for the membrane, where it will ...

  23. MXS Chaining

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    ...structures in live cells. Four individual expression constructs were created, each containing a unique fluorescent... construct. This strategy can be used to build vectors with diverse functions, and thus, is used in many...localization, as well as cellular processes like gene expression and cell migration (Sladitschek and Neveu, 2015...enhancers, polyadenylation signals, inducible gene-expression sequences, and more. By combining these components...cell cycle dynamics, titrate inducible transgene expression, or a variety of other clever applications. ...flanked with a CMV promoter (to drive high-level expression) and a polyA signal (to terminate transcription... HeLa cells show robust labeling, with strong expression and detection of each fluorophore at the respective...

  24. AAV Q&A with Tim Miles

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    ...This will be highly dependent on vector dose. The color diversity in Vector Assisted Spectral Tracing (VAST...drop in transduction coverage compared to just one vector.   Q2: Do you think it is possible to make an efficient...this scenario, you may be best served by a dual vector, split intein approach.   Q3: What’s the best method...for model organisms.    Q11: How consistent is expression is when injecting CAP-B10 or CAP-B22 in marmosets...Plasmids Adenovirus Guide Page Subscribe to Viral Vectors Blog Posts ...flexibility of AAV genome design in remarkably creative ways. The continued development of more and smaller...

  25. Five Popular Model Organisms

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    ...lab who created a set of modular vectors that can be used to optimally control gene expression in specific...spatiotemporal expression of a gene of interest. This type of control over gene expression is possible in... al., 2009). By injecting mice with retroviral vectors that harbored different mutations found in leukemia...that uses heat induction to control the temporal expression of different transgenes, the Stainer group determined...micro-injecting DNA into the worm, such as animals that express fluorescent markers (Mello et al., 1991). Genes...interference (RNAi) by feeding the worm bacteria expressing specific plasmids (L4440) that contain your gene...lifetime. Thus any genetic modification, such as expression of fluorescent protein or disruption of a gene...

  26. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

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    ...switch to be expressed. In this case, the gene encodes the LacZ𝛼 protein. Once expressed, LacZ𝛼 complexes... RNAi. Recently the Vellai lab has created an improved RNAi vector that leads to significantly stronger... RNAi is commonly achieved by feeding E. coli expressing dsRNA to C. elegans. These dsRNAs target specific...for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted...mutant animals that are genetically altered to overexpress endogenous RNAi pathways. These mutations however...non-specific nucleotides into the dsRNA leading to increased RNAi efficiency. Find the T444T plasmid here... used yeast display-based directed evolution to create APEX2. They have now applied the same directed ...

  27. Deep Mutational Scanning with One Pot Saturation Mutagenesis

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    ...codon site of interest can be cloned into an expression vector with Gibson assembly. While this is a comprehensive...relies on the generation of a plasmid library that expresses all desired variants of a protein. Applying a ...selective pressure winnows the pool down to plasmids expressing variants with optimal function. High-throughput...is generated by transforming a backbone phagemid vector into an E. coli strain that is deficient in dUTPase...2 instances where it does. 1) When the backbone vector has multiple BbvCI sites. If these sites have opposite...polymerase, 2) increased PCR extension time, 3) increased concentration of MgCl2, 4) increased concentration...PCR reaction is created by making some or all of the following modifications: 1) increased concentration...

  28. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

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    ...promoter is expressed. Cre-mediated deletion of one or several of these genes permits the expression of genes...copy of the transgene expressing either YFP or CFP, and cells that do not express Cre fluorescing with the...activate the expression of a different fluorescent protein. Therefore, each cell could express a unique combination...fluorescent protein genes, Cre recombination is required before fluorescence can occur. Cre recombinase can be...developing fish via Cre protein microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier...any cell type when crossed with mice expressing the appropriate Cre allele. The Confetti mouse contains...so that XFP are only expressed following a recombination event. Tissue-specific Cre recombination in this...

  29. Let There Be LITE Plasmids

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    ...those optogenetic controls, gene expression levels reliably increased in vitro by a factor of 10 to 20...functions, Konermann needed tools to manipulate the expression of specific genes in the brain in reversible ...Zhang showed they could package LITEs in viral vectors and genetically target them to specific cell populations...demonstrated use of the LITEs to control gene expression in mouse neurons and in the brains of living ...system could be by applying it to altering gene expression via epigenetics with the addition of histone ...different effectors. I’d encourage people to get creative with them for their own uses.” Konermann said ...found here.) So tell us, readers, how will you get creative with LITE? Reference: Silvana Konermann et al...

  30. Plasmids 101: Expressing shRNAs for RNA Interference

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    ... are transiently expressed using either AAV backbones or species-specific expression plasmids. You can...cloning strategies for constructing shRNA expression vectors. BMC Biotechnology, 6(1). https://doi.org...processing and silencing pathways. Created on BioRender.com.   Expressing shRNAs using plasmids The design...sequence. Created on BioRender.com. When it comes to the backbone, shRNAs are most often expressed constitutively...option for altering the expression of specific mRNAs. shRNAs are a quick way to create stable knockdown cells...be applied to. When it comes to altering gene expression, CRISPR technologies have gradually taken over...processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the endogenous miRNA...

  31. Single-cell tracking of lineage and identity with CellTag

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    ...were detected over a 10-week period. Their expression decreased over time but they were not silenced.    ...), now allows scientists to identify the gene expression patterns unique to the few cells that successfully...Unfortunately scRNA-seq only captures the gene expression of cells at a single time point, making it difficult...difficult to investigate how early expression patterns influence reprogramming outcomes. CellTag from ...single-cell level, thus linking ancestral gene expression patterns with reprogramming outcomes (Biddy et...MEFs were reprogrammed into iEPs by retroviral expression of transcription factors Foxa1 and Hnf4α, a process...lineage trees and comparing them with the gene expression changes of more than 100,000 cells undergoing...

  32. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

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    ...working on developing new surface display vectors to improve expression and trafficking of these biosensors....gray (PAG), with red neurons expressing tdTomato and green neurons expressing cFos. Image from Elizabeth...B) CR-Cre mice demonstrate pain response due to blue light stimulation of dual virus-expressing CR neurons...verification of the dual virus expression in CR neurons of the spinal cord. Created with BioRender.com. Image... Figure 2: Micrographs of the biosensor expressed in the human cell line, Hep3B. From left to right...opioid withdrawal with tdTomato and assessed cFos expression. She will explore the connectivity patterns of...chemogenetics to study the activity of parvalbumin (PV)-expressing neurons, neurons that the lab previously found...

  33. Plasmids 101: Gene, enhancer, and promoter trapping

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    ...fundamentally the reporter construct monitors the expression of a nearby gene. The reporter gene serves as...as a proxy for the endogenous gene – its expression levels (high/low) and patterns (tissue/timing) can...that the gene which GFP was monitoring was being expressed in the same pattern. These systems suggest the...function of a gene. For example, they eliminate the expression of the gene (by CRISPR knockout, shRNA depletion...Tissue-specific genes – the genes of interest may not be expressed in the tissue/cell line you are working with. ...the gene as a result but will still allow for expression pattern monitoring. Enhancer traps Like promoter...trapping of endogenous enhancers to stimulate the expression of the reporter. The system is made up of a reporter...

  34. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    ... how Cre-lox recombination works. Advances in Cre-lox recombination have included inducible Cre, such...membrane tethered cyan). Without Cre recombination, RFP is expressed. Cre can mediate one of two deletions... construct, OFP (orange), is expressed when Cre is absent; stochastic Cre excision results in one of three...remaining cassette can invert as long as Cre remains expressed; once Cre is removed from the system, the fluorescent...Cai et al. also created an Autobow construct containing both Cre and XFPs. Cre production drives recombination...manipulate gene expression, but other gene manipulation systems remain valuable to biology. Cre-lox recombination...temporally control gene expression, especially in in vivo models, and new Cre-lox based technologies are...

  35. Common Injection Routes in Mice

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    ...Genome Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati, P.,...be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must go through ...choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal injections...for the delivery of various therapies and viral vectors. This injection sounds very disruptive, but surprisingly... This is most often used for delivery of viral vectors, though drug treatments are also possible. One ...representation of common injection routes in mice. Image created with biorender.com.   Needle techniques Intravenous... often used when injecting a cell suspension to create xenograft models. Restraint: back scruffing or ...

  36. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

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    ...cells to express Cas9. In these particular experiments, we generated A549 cells stably expressing Cas9 via...wildly variable Cas9 expression (Figure 1). This is a problem because Cas9 expression affects growth rate...variable Cas9-expression was across the different cells (Figure 3).   Figure 2: Cas9 expression in monoclonal...Checking Cas9 expression Overview: By Western blotting (Figure 2, pool), I knew the cells expressed Cas9, but...thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low (even in...against GFP expression, but I didn’t. 2. In the original publication they see GFP expression in about 20%... MOIs. 2. Create a monoclonal Cas9-expression cell line. Especially if you’re doing a screen. I made a...

  37. Hot Plasmids - October 2022

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    ...constructs are currently available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and you can also find...B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential...voltage traces of action potentials in a HcKCR1-expressing mouse neuron without (top) and with (bottom) ... blot for the myc-tagged protein GFP-smFP-myc expressed from Plasmid 98926. See the Applications section...called JEDI-2P. Using a custom, high-throughput screening platform to apply saturating mutagenesis on 21... 1: Summary of the approach and outcomes of the screen for improved genetically-encoded voltage indicators... in-vivo imaging of voltage fluctuations. Image credit: Liu Z, et al. Cell. 2022.  Find JEDI-2p viral...

  38. Hot Plasmids: Summer 2025

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    ...synapses. In addition, ATLAS itself could be expressed Cre-dependently to allow tracing exclusively from... mGold2. Top: TIRF-SIM imaging of COS-7 cells expressing LifeAct fused to mVenus, mGold2s, or mGold2t....but TnpB is small enough to be carried by viral vectors (~400 amino acids, less than a third of the size...thousands of eligible AAV plasmids to order viral vectors ready to use in your lab. Find Addgene’s AAV6 and...postsynaptic AMPA receptor and is endocytosed, the Cre recombinase payload activates a floxed reporter gene... 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.com. Steven Jacobsen and Jennifer...

  39. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    ... 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...

  40. Antibodies 101: Affinity Tags

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    ... single protein is expressed, with the intent of purification, it will be expressed along with all other...The MBP tag, although large, can actually increase expression and solubility of the tagged protein (Fox...other cellular proteins within the expression system (bacteria, yeast, mammalian, etc.). To individually...about that protein that no other protein in the expression system has. This is where affinity tags come ...peptides which do not naturally exist in most expression systems that can be added to the N or C terminus...on: The protein you are trying to purify The expression system If you want the tag to remain on your ...Ready to start tagging? We are here to help with vectors, protocols, antibodies, and more!   Resources ...
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