Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 920 results for: Gene

Showing: 41 - 80 of 920 results

  1. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ...and easily manipulate specific genes. But what if you want to study genes all across the genome? Two new...7,114 human genes. Unlike the GeCKO library, each library has been enriched for different gene targets. ...designed to target a smaller set of genes and it includes more guides per gene. The Sabatini/Lander library ...screen for genes involved in bacterial toxin resistance, identifying 4 previously unknown genes.  “With ... function of both protein-coding genes as well as non-coding genetic elements.” These lentiviral libraries...library, which they call GeCKO, targets 18,080 human genes with 64,751 unique guide sequences to enable both...including Feng Zhang used the GeCKO library to identify genes essential for cell viability in cancer and pluripotent...

  2. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ... site (TSS) of a gene of interest, it induces potent gene expression with no genetic alterations observed...efficiently transcribing the gene. This results in gene silencing, with no observed genetic alterations at the ... of modulating gene expression makes it an invaluable tool in the interrogation of gene function. However...exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast...need to generate new cell lines or transgenic animals expressing dCas9 as a means of modulating gene expression...Cas9 can be reprogrammed to serve as a regulator of gene expression. Its catalytic domains can be mutated...with a gRNA that directs them to the promoter of a gene of interest, they have been shown to dramatically...

  3. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ... transcription of a target gene, it therefore determines the timing of gene expression and largely defines...These permit exquisite control of gene expression and tracing of gene expression across development. One...of herpes simplex virus immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101...systems are genetic tools that consist of two parts, both of which must be present for a gene of interest...placing UAS upstream of a transgene permits directed expression of that gene in cells that also express...expression of reporter genes, or to combine reporter expression from one promoter with transgene or siRNA expression...together for complex genetic analysis in Drosophila or for precise control over multiple genes in synthetic systems...

  4. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ... identify genes by their similarities to genes in other species, but these studies generally leave questions... confident that the gene from the test species has a similar function to the gene in the reference species...MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes from...questions about gene function wide open. What if two seemingly similar proteins have acquired divergent...straightforward. Instructors first select a multi-gene pathway to study and a reference organism where ... to answer questions about the evolution of the genes involved in methionine synthesis (5). The budding...strains with overexpression plasmids carrying MET/Met genes from either S. pombe or S. cerevisiae. Students ...

  5. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ... applications Gene expression control Researchers are always wanting to manipulate gene expression – turn...viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA targeting...Additional Resources on the Addgene Blog AAV: A Versatile Viral tool for Gene Expression in Mammals Listen...from basic science research all the way to clinical gene therapy. But how can you use these amazing tools... knock it out, etc. It’s how we understand what genes do! Viruses have several options to allow you to... Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily, long-term, and even...engineered to introduce frameshift mutations in genes of interest, turning expression off permanently....

  6. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...expression of Gene X using shRNA expressed from a plasmid   Figure 1: Expression Level of Gene X The results... level of Gene X in cells treated with Plasmid A is similar to the expression level of Gene X in the untreated...backbone itself on gene expression in your target cells. In experiments employing gene targeting or genome...trying to repress a gene, your control might be a system where expression of that gene is knocked out completely... shRNA targeting human Gene X in Plasmid A is the decreased expression of Gene X. Ergo the positive control...targeting human Gene X. The non-targeting control also assesses any effects of the general introduction ... wherein Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected into human cells...

  7. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...articles, genes, and blog posts are not comprehensive, so just because you do not see a name or gene on this...plasmids containing these genes. As with the PIs above, many of these genes were discussed at the 2018 Keystone...HIST2H2BE RBX1 STAG2 Xist Your favorite gene! SWI/SNF Genes:       BRG1(SMARCA4) BAF170 (SMARCC2...You!     Chromatin-related genes with plasmids available from Addgene The links below pull up lists...containing the indicated gene, and you can subscribe to these pages to learn when Addgene receives new plasmids... In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1 CREBBP CUL1 H2AFX ...Interactions: Using CRISPR-dCas9 to Study Gene Regulation CRISPR 101: Epigenetics and Editing the Epigenome CUT&RUN...

  8. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ...Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used to study gene expression...for visualizing gene expression in vivo.  But what ultrasound lacked was reporter genes that could produce...Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when expressed, enable cells...vesicles gene clusters from a diverse set of gas vesicle-expressing species. The gas vesicle genes from Serratia...structural gene gvpA marked with EBFP2 and the second cassette contains the P2A-linked accessory genes gvpNJKFGWV...first-generation mammalian ARGs (mARGs) enabled ultrasound visualization of in situ mammalian gene expression...hypothetical protein, improving expression, and named this gene set bARGSer, for bacterial ARG from Serratia sp....

  9. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    Published
    June 11, 2020, 1:34 p.m.
    ...find AAV tools for optogenetics and chemogenetics on the Addgene website. Optogenetics also requires that...molecule activation (chemogenetics) or light (optogenetics).   Timing Optogenetics offers a high degree...limitations.  The four main factors in which chemogenetics and optogenetics differ are: Timing Targeted manipulation...schematic showing the differences in chemogenetics and optogenetics in a mouse experiment. Viral delivery...faster.  Chemogenetics does not offer the same level of control over timing. Because chemogenetics relies...required. Targeted manipulation Both optogenetic and chemogenetic tools can have their expression targeted...stimulation is gradual. Invasiveness Both optogenetic and chemogenetic tools need to be delivered to cells,...

  10. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...resistance gene, NPTII, facilitates phosphorylation of kan, which inactivates it. The NPTII gene confers ...death by intercalating into DNA and generating DNA breaks. The Sh ble gene provides zeocin resistance by expressing...the Sh ble gene does not fully suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your.... But does it matter which antibiotic resistance gene you select? In this blog, we’ll break down the mechanism...action of the most common antibiotic resistance genes and which applications are best (or worst) for each...plasmid, isolate individual plasmids, or express a gene contained in a plasmid. During the propagation and...

  11. Antibodies 101: Isotypes

    Type
    Blog Post
    Published
    Oct. 19, 2021, 1:15 p.m.
    ...constant genes μ and ẟ. These are the first two genes in the heavy-chain constant domain gene cluster....constant domain genes, placing the variable domain genes next to a different constant domain gene. This process...Heavy chain constant genes are clustered together downstream of the variable region gene in this order: μ,...constant domain gene. For example if a B cell switches to producing IgE antibodies, the gene segments for...nomenclature. Antibody heavy chain proteins as well as the genes that encode those proteins are designated by the...μ, δ, γ, ε, and α. An antibody variable gene is initially expressed with the IgM and IgD constant domains...domains because they are the first constant domain genes in the cluster. However, when B cells switch to ...

  12. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    Published
    July 28, 2017, 1:30 p.m.
    ...The white gene is most often used, but other selection markers include vermillion (another gene controlling...as species of gene. Find Gal4 and UAS plasmids at Addgene Choosing a vector to generate a transgenic fly...described the Gal4/UAS system used by geneticists to study gene function. In this final installment, I...you’d like to use to generate your new fly line: 1. Where would you like the gene to be incorporated into...spot flies with the appropriate transgene. One such marker is the white gene, which restores eye color to...engineer new transgenic flies to study your favorite gene (YFG).  Sometimes, you want to use the Gal4/UAS...fairly straightforward. Basically, you place your gene of interest in a plasmid and get the vector incorporated...

  13. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    Published
    June 19, 2018, 1:38 p.m.
    ... Acoustic reporter genes (ARGs) Acoustic Reporter Genes (ARGs) are engineered gene clusters that encode...for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference between...Resources on the Addgene Blog Learn more about in vivo imaging tools The luciferase reporter gene The green ...green fluorescent protein reporter gene Resources on Addgene.org Find plasmids for in vivo imaging with ... reporters has its drawbacks. Acoustic reporter genes (ARGs) overcome these limitations by using gas vesicle...and high spatial resolution. By piecing together genes from gas vesicle operons from two different bacteria...their ultrasound signal. Using acoustic reporter genes for in vivo imaging Following in vitro characterization...

  14. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...numerous applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac...levels of your gene of interest, as needed. Controlling the expression levels of your gene may be particularly...Transposagen.   How does piggyBac™ enhance gene editing? Gene editing can now be accomplished in a more... How does the piggyBac™ gene editing system compare with other methods of gene editing? There are primarily...Decoding the Epigenetic Heterogeneity of Human Pluripotent Stem Cells with Seamless Gene Editing.,” Methods...remain, such as making gene edits completely seamless (i.e., so no other genetic alterations have occurred...cell lines and genetically-modified cell lines. Furthermore, the use of piggyBac™ for gene editing allows...

  15. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ...any CRISPR-based gene editors, for correcting mutations or installing functional genes to study and potentially... number of genes with a single viral vector.   Fig. 1: Schematic of multiplex gene editing with...enable multiplex gene editing in a variety of cells and organisms. However, lengthy genetic payloads and ...demonstrating simultaneous editing of genes against multiple genetic diseases, such as muscular dystrophies...limited the scalability and efficiency of multiplex gene editing. Moreover, existing multiplex strategies...single array, has been a top choice for multiplex gene knockout or regulations, it lacks proper functions...derived tools are leading the edge of the precision gene editing. CRISPR-Cas9, the key component to enable...

  16. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...Adenovirus Vectors for Gene Therapy, Vaccination and Cancer Gene Therapy. Curr Gene Ther, 13(6), 421–433...compartment, gene expression, and protein translation. Its genome consists of “early” (E) and “late” genes, and...with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions are rescued by helper plasmids... be expressed, since the early genes are absent, and the late genes (and their structural proteins, which... does not require de novo viral gene expression: Implications for gene therapy. Proc Natl Acad Sci U S...house a recombinant sequence (transgene) instead.    Although the late genes are usually present in the ...and E3 gene complementation. In target cells that are infected by the vector, only the transgene will be...

  17. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...hosts through horizontal (or “lateral”) gene transfer. Horizontal gene transfer can occur naturally through...carrying genes for bacterial toxins like colicin typically also carry the necessary resistance genes to help...Additional Resources on the Addgene Blog Plasmids 101: Antibiotic Resistance Genes Antibiotic Resistance: ...cell to another). Bacteria need special fertility genes to be capable of conjugation, and these are encoded...there is evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001...organisms to thrive in competitive environments. The genes encoding these functions form the basis of many ... Resistance (R) plasmids: microbial defense Genes allowing bacteria to survive toxins are the clearest...

  18. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    Published
    May 7, 2019, 12:39 p.m.
    ... several genes spanning 33 cancer types. The types of mutations they identified included gene amplifications... plasmids each carrying a single gene with a defined mutation or gene fusion. Plasmids carrying the corresponding...how a given gene is involved in a complex pathway. Some of the most heavily-studied genes in cancer research...help cancer researchers studying an assortment of genes. Using patient data to identify mutations of unknown...corresponding wild-type genes were constructed as well. They called this library the CTD2 FASMIC Plasmid...thoroughly investigate any of the dozens of wild-type genes and their associated mutants in other model systems... Yiu Huen, et al. "Functional annotation of rare gene aberration drivers of pancreatic cancer." Nature...

  19. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...spatiotemporal expression of a gene of interest. This type of control over gene expression is possible in ...diseases that are caused by a specific gene. For example, several genes mutated in human cancers have a homolog... They share about 70% of their genes with humans and 85% of human genes associated with a disease have... share many genes with humans, are easily maintained in the lab, and have short generation times that ...Thus any genetic modification, such as expression of fluorescent protein or disruption of a gene, can be...Refinement of tools for targeted gene expression in Drosophila." Genetics 186.2 (2010): 735-755. PubMed ... of Drosophila research. Morgan discovered that genes were found within chromosomes using the fruit fly...

  20. Plasmids 101: Modular Cloning

    Type
    Blog Post
    Published
    March 12, 2024, 1:15 p.m.
    ...platform. Gene Stacking As the name implies, gene stacking is the use of a combination of multiple genes to ...regulate the transcription of specific genes. What if one of those genes coded for a protein that regulated...combinations of different candidate genes at once to identify the best cocktail of genes when the relationship between...transcription of another gene? Now you have a genetic circuit! Genetic circuits are multigene systems whose protein...characterized activities with predictable behaviors. While a gene is the most obvious example of an individual part...transcribed together. They can express one or more gene(s), and typically begin with a promoter and end ...could be comprised of just three parts: promoter, gene, and terminator.     Fig 1. MoClo enables...

  21. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...Positive-selection vectors using the F plasmid ccdB killer gene”. Gene. 1994 Oct 11;148(1):71-4. PubMed PMID: 7926841... experiments, and here we will focus on a unique gene that makes the popular GatewayTM method possible...accomplish this? Let’s start with a brief history of the gene and how molecular biologists have harnessed it to...cloning technology. CcdB: A potent toxin... The ccdB gene, located on the F sex factor plasmid of E. coli,...and ultimately causing cell death. ccdA, another gene found in the ccd operon, codes for the antitoxin...vectors, called pKIL18 and pKIL19, contained the ccdB gene in frame with an MCS. E. coli that were transformed...transformed with the empty vectors expressed the ccdB gene and were therefore unable to propagate because CcdA...

  22. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...strategy is to knockdown a gene with RNAi. Here, a short sequence targeting the gene is cloned into a viral...whether it is an overexpression gene, downregulation via RNAi or gene editing/deletion using CRISPR. In...conjunction with a genetic manipulation so that behavior can be linked to a specific gene or brain region...viral vector should be used? Modifying gene expression Generally speaking, one may want to achieve either... application.  Genetic manipulation can also reduce expression levels of certain genes. A popular method...can now precisely control where and when certain genes are expressed to investigate their role in a specific...specific brain region. One option is to overexpress a gene that alters neuronal function directly such as receptor...

  23. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    Published
    April 28, 2016, 2:30 p.m.
    ... the gene you would like to rescue with in the antisense orientation (let’s call the gene “GeneR”). The...rescue of a gene knockout: To test if the phenotype you are observing is really due to a single gene deleted...endogenous gene. Spatio-temporal expression of optogenetic reporters: Since most optogenetic reporters...transduced reporter gene. Generating a FLEx switch to control expression of an optogenetic reporter would ... were designed as a genetic tool for researchers to conditionally manipulate gene expression in vivo using...conditionally turn off one gene and activate another. In the field of mouse genetics, FLEx is currently being... vivo conditional gene rescue experiments, as well as for improvement of optogenetic adeno-associated ...

  24. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...Entrez Gene matches to your insert. Select the appropriate gene if it appears in the Entrez Gene list, ...correct Entrez Gene does not appear in the auto-populated list, try entering the gene's Official Full ...known as" aliases in the Gene/Insert box. Once you have selected the appropriate gene from the list, save ...bacterial strain(s) through Addgene. Description – please indicate the gene cloned into your plasmid, any... for the gene or insert. Step 4: Growth and Distribution Please indicate which strain Addgene should propagate... Encodes insert(s) A plasmid with one or more genes, protein coding sequences, or other elements inserted...Empty backbone A plasmid designed for cloning a gene/insert into it. Empty backbones often contain epitope...

  25. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...recombination; ccdB gene for negative selection Used to clone attB-flanked genes of interest to generate entry clones...for recombination; ccdB gene for negative selection; elements to express the gene of interest in the appropriate...for gene expression.     The LR Reaction takes place between the attL sites of the generated entry...attB sites is generated. As in the BP reaction, a DNA fragment containing the ccdB gene is excised from... cloning to generate our desired constructs: lentiviral expression of the human KRAS gene in mammalian...   Pro Tip: Addgene also has ready-made entry clones available for many popular genes, including Hs....our website to search for your favorite gene! STEP 2: Generate an Expression Clone When making the expression...

  26. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...activation sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless...promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...reporter gene (3, 4), there are now many variations, including many that use auxotrophic rescue genes as reporters...Shapira S.K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in Escherichia coli and ...hybrid system (also called the interaction trap), genes encoding proteins of interest(POI’s) are fused to... in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing the target... the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI’s interact...

  27. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ...approaches increase the size of the gene delivered by splitting the gene into two pieces: part A and part...packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice. ...how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces. ...acceptor sequence, the 3’ half of the transgene. The full length gene is reconstituted by either the overlapping...Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human Gene Therapy...Chamberlain JS (2011) Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination ...retinal gene therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check...

  28. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...read-through transcripts of the effector gene. Effector genes, any gene that you want to control the expression...chemical inducers of gene expression (del Valle Rodríguez et al., 2011).  Genetics of SPARC Figure...Learn more about controlling gene expression in flies Resources on Addgene.org Find fluorescent protein...truncated attP sites to control recombination of the gene construct. A predictable number of cells will express...binds to the UAS binding motif, driving downstream gene expression. attP and attB, recombination sites. ...side of a stop cassette, upstream of an effector gene. In cells where the stop cassette is removed during...expression of either a stop cassette or an effector gene. HDVR (2X Hepatitis Delta Virus Ribozyme), a self-cleaving...

  29. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...typically include a gene that is advantageous for survival, such as an antibiotic resistance gene. In contrast...the target gene. Vital component for expression vectors: determines which cell types the gene is expressed...scientists use plasmids? Generally, scientists use plasmids to manipulate gene expression in target cells...replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing bacteria...MCS is often downstream from a promoter. Insert Gene, promoter or other DNA fragment cloned into the ...obtained. Selectable Marker The antibiotic resistance gene allows for selection in bacteria. However, many ...referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may utilize...

  30. Donations from Addgene to Yield Answers for Rare Disease Researchers

    Type
    Blog Post
    Published
    Sept. 15, 2015, 2:30 p.m.
    ...CRISPR-Cas9 constructs from Addgene to gene-edit mutations into XP genes and other genes involved in photosensitivity...researchers identified the gene responsible for the disease as ATP1A3. The gene encodes a sodium-potassium...Syndrome, a devastating genetic disease caused by mutations in the OCRL1 gene that leads to the early ...studies on OCRL1 already underway in his lab. The gene appears to play a role in many cell processes involving... learned there was a disease associated with the gene, it piqued their interest. They obtained cells from...McSweeney says they now think that mutations in the gene may prevent neurons from returning to basal conditions...Wattanasirichaigoon carried different mutations in a gene called SOX10. To understand the functional role ...

  31. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...between MARCH5 and WSB2 genes using Cas12a’s ability to knock out multiple genes in parallel. Lastly, this...to induce high levels of gene activation for both reporter and endogenous genes. It is highly specific ...cellular targets Optimized genetic screening technique using Cas12a for multiplexed gene perturbation The Penn...10846-1.   Optimized genetic screening technique using Cas12a for multiplexed gene perturbation by: Gabrielle...many CRISPR-based genetic screening methods have primarily used Cas9 to knock out genes, researchers at ...multiple genes within a single cassette provides increased simplicity for combinatorial genetic screens...frankenbodies to amplify the signal.  Liu et al. Genes Cells. 2021. https://doi.org/10.1111/gtc.12893, ...

  32. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    Published
    Oct. 19, 2023, 1:15 p.m.
    ... with ITRs that flank the gene cassette, which contains a promoter, the gene of interest, and a polyA ... DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs...modifications can drastically affect gene transfer efficiency and transgene expression (Zhou et al. 2017). ...Terminal Repeat Sequence Role in Vector Transgene Expression. Hum Gene Ther, 31(3–4), 151–162. https://doi.org...biological tool that allow scientists to express a gene of interest in a cell or animal model. AAV plasmids...ITRs, and the ITRs are retained with the gene in a recombinant AAV (rAAV) vector (Figure 1). Everything... virus. In an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic...

  33. What's New in CRISPR - March 2020

    Type
    Blog Post
    Published
    March 10, 2020, 1:15 p.m.
    ...demethylation Optogenetic repressors of gene expression in yeast Megan McClean’s lab generated an optogenetic...2019. Fusing epigenetic modifiers such as TET1 with inactive Cas enzymes can alter gene expression via...optogenetic toolkit that controls repression of gene expression in Saccharomyces cerevisiae. The system consists... enters the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the NLS is inaccessible...dilutions of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from...takes several weeks to delete or deplete individual genes. Michael Laub’s lab developed a CRISPRi system to...downregulate expression of specific Caulobacter genes using a catalytically dead Cas9 (dCas9) derived ...

  34. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ...you are trying to do: gene knockout, a specific base edit, or modulation of gene expression. Location ...Created with BioRender.com. The hammer: Gene knockout by NHEJ Gene knockout with CRISPR technology is usually...multiple sequences targeting a gene to score in order to call that gene a hit.  This is an exciting time...optimize numerous parameters to generate the desired edit. The wrench: Gene activation and inhibition by ...edits and to perform functional screens to identify genes involved in a phenotype of interest. This blog post...indels, it's not so important what location in the gene you target, but it is important that your gRNA sequence...creating a non-functional allele. For a 1 kilobase gene, since potential target sites occur ~1 in every ...

  35. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    Published
    July 28, 2016, 2:30 p.m.
    ...possible to build gene regulatory RSMs (GRSMs, the subset of RSMs that regulate gene expression) that ...large serine recombinase genes downstream of inducible promoters: the BxbI gene downstream of the anhydrotetracycline...the TP901 gene downstream of the arabinose (Ara)-inducible promoter (PBAD), and the A118 gene downstream...register to execute predictable gene regulation programs that specify which genes should be expressed or not... same gene regulation program. We recommend testing multiple register designs for the same gene regulation... paper. The paper also demonstrates that genetic parts (genes, terminators, and promoters) may be interweaved...promising strategy for linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange...

  36. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    Published
    July 12, 2016, 2:30 p.m.
    ... so genes. It would be great if we could just go in and quickly manipulate individual mouse genes and ...insertion of a reporter gene, or replacement of the murine sequence with a human gene. Advantages of using...conventional gene targeting. CRISPR tools were recently used to introduce a 25 kb human gene into the mouse...conventional gene targeting. It also costs less. With CRISPR, it may cost about $5,000 to generate founder ...for Tips on CRISPR Gene Editing in Mice Disadvantages of using CRISPR for generating mouse models Although...Jaenisch R (2013) One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated ...Haoyi Wang. CRISPR/Cas9 is revolutionizing the mouse gene-targeting field. Mice have long been extremely useful...

  37. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ...References on Addgene.org Check out Addgene's Chemogenetics Guide Find fluorescent proteins at Addgene ...on the Addgene blog Browse Fluorescent 101 blog posts Read about chemogenetics vs. optogenetics Find more...Emory University School of Medicine. Optogenetics and chemogenetics are powerful tools to modulate the ...neurons and other brain cells. Since the opsins or chemogenetic receptors used in these techniques cannot be...versatile method to identify the expression of transgenes of interest. Furthermore, fluorophore variants... important influence on the expression of the transgene of interest and its localization in the membrane... tag protein can impact the expression of the transgene of interest. In our studies, mCherry interfered...

  38. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...is inserted at the 5’ end of a gene, the SEC separates the targeted gene from its promoter, creating a ...the targeted gene. After initial testing, Dickinson et al. tested this system across 7 genes in parallel...animals per gene, they obtained the desired insertions on the first attempt for 6/7 genes. The data are...hands-on time. Creating a modular, selection-based gene tagging approach Multiple features of C. elegans...uses a selection cassette containing genes for hygromycin resistance and the Roller movement phenotype. To...design and clone 500-700 bp homology arms for your gene of interest into the FP-SEC vector using Gibson ...loss-of-function allele as well as a promoter-FP fusion for the gene of interest. L1/L2 larvae from these strains are...

  39. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...sphaeroides Cas9, dCas9 Gene editing Zhou et al., 2018   pdDronpa Cas9, dCas9 Gene editing, transcription...showing that this genetically encoded, photocaged Cas9 system could silence endogenous gene expression - demonstrating...photocaged-gRNA Gene editing For more details, check out this comprehensive review comparing optogenetic CRISPR...a system that would use light to turn on and off gene expression while imparting spatiotemporal control...and move the activator into position to activate gene transcription. The researchers tested a variety ...researchers showed that expression of the human ASCL1 gene could be spatially controlled in a reversible and... to develop an optimized photoactivatable CRISPR gene activation system, but settled on a different optimal...

  40. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...library that has 10 gRNAs per gene and targets all ~20,500 protein-coding genes. Unique, non-overlapping sites... identified ricin toxicity genes. The screen also discovered several genes previously not associated with...p-values of known ricin regulation genes compared to results generated using non-targeting guides as controls...-throughput systematic approach for identifying genes involved in a biological process. These screens ...toxicity screen since essential and non-essential genes for both pathways had previously been identified...identifying >88% of previously identified essential genes while previous Cas9 and shRNA library screens only...only identified  60% of the essential genes. The ricin toxicity screen had a 10% false discovery rate ...
Showing: 41 - 80 of 920 results