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We narrowed to 920 results for: Gene

Showing: 161 - 200 of 920 results

  1. The Importance of High Titer for AAV Transductions

    Type
    Blog Post
    Published
    March 1, 2018, 2:11 p.m.
    ...Versatile Viral Tool for Gene Expression in Mammals Additional Resources on Addgene.org Find Ready-to-Use ...the cell on their way to the nucleus where their gene cargoes can be expressed. The bottom line is that...successful transduction. In another example, neuronal gene expression by an anterograde AAV was shown to drop...cell, you may not be able to detect AAV-mediated gene expression in a target cell. Practical considerations...experimental setup. Some experiments require higher gene expression and therefore higher titer than others... to such a high titer when looking for effective gene expression. How do you achieve high titers in the...therapy : the journal of the American Society of Gene Therapy. 2008;16(7):1189-1199. doi:10.1038/mt.2008.103...

  2. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...EGFP can be used as positive controls for gene editing or gene activation, while the BRAF and MAP3K4 gRNAs...provide the Brunello (targeting human genes) and Brie (targeting mouse genes) pooled libraries from the labs...editing. You can also enhance expression of your gene of interest with a variety of Cas9 activators available...pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular processes...80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to both perform...and member of the Addgene Advisory Board) says: "When you're setting up to do a genetic screen, you need... are members of the Addgene Advisory Board. Additional Resources on the Addgene Blog Get the Most from...

  3. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...alternative method for generating transgenic mice strains and for non-invasive in vivo gene editing. The bottom...repression, transgenic mouse line creation, and in vivo gene editing of mice, with the potential to be used for... Primary cells are often refractory to gene delivery methods, but the researchers was able to edit hiPSCs... stimulation, respectively. Primary cell type Gene targeted by guide Editing efficiency hiPSCs  EMX1...cells when using a gRNA targeting the Fto gene, while editing was not detected in cells electroporated with... used Nanoblades to disrupt the tyrosinase (Tyr) gene in mouse zygotes, which results in albinism in mice....  Nanoblades can also be used to non-invasively gene edit the liver of adult mice. When mice were IV ...

  4. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    Published
    July 18, 2019, 12:55 p.m.
    ...toolkits available at Addgene.  C. elegans open resources The C. elegans Neuronal Gene Expression Map & Network...expression of your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene, we have the entire...-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans." Genetics 202.4 (2016): 1277-1288... The CeNGEN goal is to establish a comprehensive gene expression atlas of every C.elegans  neuron. Scientists... C. elegans embryo when all essential embryonic genes are individually depleted via RNA interference (...based bipartite expression system that can regulate gene expression in C.elegans. In this system a promoter...that can bind to activation sequences upstream of genes to stimulate expression (Wang et al., 2016). In ...

  5. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...discussed the promoter's role in the initiation step of gene transcription; today we'll provide an overview on...define the end of a transcriptional unit (such as a gene) and initiate the process of releasing the newly...machinery. Terminators are found downstream of the gene to be transcribed, and typically occur directly ...studies focus on promoter strength as a determinant of gene expression levels, the terminator also plays an ... to variability in RNA half-life, and ultimately gene-expression. Polyadenylation is the post-transcriptional... in a Promoter-Dependent Manner. Hum Gene Ther. 2008. PMID: 18627247 Zufferey R, Donello JE, Trono D, ...

  6. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...Repair is Used to Edit Genes Read How Non-Homologous End Joining is Used to Delete Genes Additional Resources... CRISPR gene therapy, it’s important to realize that DMD is a simpler case than other genetic diseases...there is still no good treatment. The dystrophin gene is very large (79 exons), but much of the sequence...Dystrophin’s size makes it difficult to deliver via gene therapy, so researchers have set their sights on... cells expressed functional dystrophin. Editing genes in vivo is of course much more difficult than in...cells, where precision editing of the dystrophin gene would take place, with minimal risk of off-target...clinically with CRISPR. Many obstacles remain for CRISPR gene editing, but given the speed at which the technology...

  7. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ...through this challenging sequence. Transgene The transgene is the gene that will be delivered by the AAV..., or your gene of interest. Promoter The promoter drives the expression of the AAV transgene. On a plasmid...plasmid which contains the structural and packaging genes, the adenoviral helper plasmid which contains the...plasmid map, promoters are upstream or 5’ of the gene they control. Promoters can be organism or tissue...for the successful and specific expression of your gene of interest. References Haery L, Deverman BE, ...Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods 30:206–213 . https://doi.org/10.1089...control transgene expression. Lox sites are directional 34 bp sequences that can flank the transgene and ...

  8. RUBY-Red Siliques

    Type
    Blog Post
    Published
    March 1, 2022, 2:15 p.m.
    ...fluorescent proteins. Selection using resistance genes usually involves killing the untransformed seedlings...individual. For example, with CRISPR-Cas9 gene editing, transformation may be identified by resistance ...continued production of new alleles or off-target gene editing. Antibiotic selection would only then kill...requires specialized equipment. Reporters for studying gene expression have similar drawbacks, often requiring...Res. (2020).   I can do this by expressing a gene that rescues the mutations, using RUBY as a selection.... et al. A reporter for noninvasively monitoring gene expression and plant transformation. Hortic Res ...presence of a transgene instead, which can’t be done if your selection method kills the transgene free individual...

  9. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...repression of your target gene, activation and repression of different genes in a single cell, and localization...replication and transcription as well as reporter genes. To learn how you can use minigenomes in your research... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (... (AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are...localization of a fluorescent signal to your target gene. Read this post to find out more about how the Casilio...are widely used by the optogenetics and chemogenetics communities to activate neurons and neuronal pathways...guest writers. Your contributions make Addgene and the Addgene Blog useful resources for the biological...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...cross-cassette interactions in gene expression, and to generate multi-gene knockouts. Mammalian and fly ... to generate anything from individual transcription units (TUs) used to express your favorite gene-of-...While a generic MoClo system for any given species can be repurposed for CRISPR applications (gene insert...DNA assembly tool used to build single- and multi-gene constructs for a variety of applications. MoClo ...several additional capabilities, including conditional gene expression, multiplexed CRISPR, and chromosomal ...). Those looking to express high levels of their gene of interest in yeast should also take a look at ...elements for S. cerevisiae chromosomal integration and gene expression, the system includes a toolkit to express...

  11. Top Requested AAV of 2017: pmSyn1-EBFP-CRE

    Type
    Blog Post
    Published
    Jan. 17, 2018, 2:57 p.m.
    ...Cre-dependent gene of interest in neurons. This ability to be paired with any Cre-controlled gene of interest...neuroscientists because they efficiently deliver genes to various brain regions. You can partially control...occasionally you may want to further control where your gene of interest is expressed.  A cell-type specific ...and what do they mean? AAV: A Versatile Tool for Gene Expression in Mammals Subscribe to Viral Vector ...! To find more viral vectors available through Addgene, be sure to check out our viral service page and...Vector Blog Posts Additional Resources on Addgene.org Check out our Viral Service Page Browse Additional ...

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

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    ... allowing for the gene on the toehold switch to be expressed. In this case, the gene encodes the LacZ𝛼...interfere with other genetic pathways making it difficult to link a phenotype to the gene being targeted via...need for mutant genetic backgrounds. The standard RNAi vector, L4440, places a target gene between two inverted..., highly efficient RNAi construct for C. elegans gene knockdown Article contributed by Alyssa Cecchetelli...extensively used in C. elegans research to study gene function. RNAi is commonly achieved by feeding E... from each promoter progresses past the specific gene producing long unspecific RNA fragments that are... most scientists use mutant animals that are genetically altered to overexpress endogenous RNAi pathways...

  13. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...stably integrate a reporter gene that contains a puromycin resistance gene followed by the target site...that targets your desired gene (whose sequence is also found in the reporter gene). A control RFP expression...donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP. This visual read ... express RFP while cells containing the reporter gene that was cleaved by Cas9 and repaired via HDR using...unedited cells. In an experiment that targeted the gene NFE2L2, the Madison lab found that H2B-GFP expressing...desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent protein...stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems...

  14. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...inducible yet permanent gene excision. Essentially, LoxP sites are knocked in flanking a gene of interest (or ... the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a frameshift...? Is the regulation of an exogenously introduced gene or over the native locus? Let’s explore each of...Cas9 fused to a transcriptional repressor to your gene of interest. It will inhibit transcription instead...cinch. Stably introducing the coding region of a gene (cDNA) can be done by either plasmid or virus. A...Cas9. The system is led, via a custom gRNA, to your gene of interest to temporarily increase expression. ...of action Check out Addgene's Lentiviral Tet-on and AAVS1 targeted Tet-on transgene vector. Degron tags...

  15. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ...all known genes. This is a perfect example of an unbiased, reverse screen – all known genes are assayed...of knocking out the genes of interest (or the whole genome), they knock down gene expression temporarily... a biological process. Players can be proteins, genes, enzymes, drugs, or anything else that plays a role...where there is removal of known factors (such as a genes) and the loss of those factors is assessed. The ...especially useful if you are interested in studying genes whose knockout would be lethal to cells and thus...The two most common methods of manipulation are genetic knockout mutations and treatment with small molecules...selection would work is if you are trying to identify genetic conditions which convey resistance to a drug. You...

  16. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...functional groups of genes involved in the execution of a specific regulatory regime (such as genes involved in...consortium-generated datasets with a systematic effort to compile NR- and coregulator-related global gene profiling...remain unknown. Together, NRs govern expression of genes involved in a broad range of reproductive, developmental...efficient and timely activation or repression of gene expression (see www.nursa.org for a curated list... NRs and coregulators, and although the reporter gene has over the years evolved from chloramphenicol ...O'Malley, B. W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators...funding model in nuclear receptor research, and has generated a set of comprehensive datasets that collectively...

  17. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...specific genes involved. Downregulation of tumor suppressors and upregulation of proto-oncogenes ­­­can ...click on a gene of interest will direct the investigator to a list of all available Addgene plasmids he... group of genes/proteins that, when expressed, normally encourage cell survival, growth and proliferation... the health of an organism that both classes of genes function normally to maintain balance – encouraging...Technologies, Carlsbad, CA) that represents 180 genes from the Ras signaling pathway. Its major appeal...of vectors to study the function of any of these genes through several different applications such as fluorescent... in the development of cancer. With hundreds of genes involved in the Ras pathway alone and thousands ...

  18. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh (Precise Integration into Target ...than that observed with NHEJ. PITCh: Using MMEJ for gene knock-in Building on the lab’s previous work, Sakuma... should target near the last coding exon of your gene of interest. For ideal use, this gRNA should be ... is if the GFP-Puro will alter expression of the gene it follows. For increased versatility, it would ...would be advantageous to adapt PITCh to insert genes into AAVS1, the “safe harbor locus” of the human genome...another strategy researchers can exploit for CRISPR gene editing. In organisms where HDR is downregulated...Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh...

  19. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ... and the gene that is responsible is understood. It’s possible to actually silence this gene with RNAi...only 1,700 genes total). Hydra (Celina Juliano) This is a known stem cell and regeneration model. Little... human interaction for all of us studying human genes in test tubes and gels). As a freshman in college... I studied development and evolution of histone genes in Strongylocentrotus purpuratus, the spiny purple...direct tests of cause and effect hypotheses between gene function and phenotypes in diverse plants, animals...frog cell lines to develop robust methods for frog gene editing. They edit in a cell line, then take the...nonetheless - the first example was of a pigmentation gene.   Genus Microtus (Zoe Donaldson) Voles have fascinating...

  20. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    Published
    July 17, 2018, 12:23 p.m.
    ...courses on genetic manipulation more hands on While teaching courses about gene therapy and genetic manipulation...that similar types of thinking go into developing gene therapy strategies for various diseases. That’s ...Inserts, Tags, Origins of Replication, Resistance gene and Selection Markers) and one “Transfer Plasmid...had to pick the best strategy and propose a final gene therapy approach for the disease to be openly discussed...impression was that they had clearer ideas about gene therapy in the following lessons. And let me tell...teach gamma-retrovirus or lentivirus-based ex-vivo gene therapy’ strategies to undergraduates, however you...problem is…how can you make a “hands-on” lesson on genetic manipulation of human stem cells when students ...

  21. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...systems allow researchers to understand bacterial gene clusters that produce useful natural materials, ...capable of robust expression of high G+C content genes, as validated through monitoring the expression ...human mRNA binding proteins with 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode...pooled library from Junjie Chen's lab targets human genes known or suspected to be involved in DNA damage ...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons in ...library uses CRISPR cytosine base editors to disrupt gene function by targeting start codons, splice acceptor...find all of the CRISPR plasmids available from Addgene, head over to our CRISPR Plasmids and Resources...

  22. An Addgene Summer Internship

    Type
    Blog Post
    Published
    Aug. 8, 2024, 7:22 p.m.
    ...choose what gene to focus on. The gene choice had restrictions. It couldn’t be a Covid gene, a toxin, or...Syntaxin3 gene was the winner. We used the next 4-5 weeks to work on cloning the Syntaxin3 gene in a pLenti...after discussion, we sent it for sequencing. The gene was successfully sequenced, confirming that we had...with Amy C., Addgene’s intern coordinator shortly after hearing her talk about Addgene’s internship opportunity... past six weeks. Through Addgene, I learned the basics of lab safety, general work skills — such as using...environment changes them. Addgene gave me the chance to become a real life scientist. Addgene gave me the chance...knowledge more than ever before. Before joining Addgene, I had spent most of my junior year stressed out...

  23. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ... plasmids to generate deletions of candidate genes in euploid iPSCs. “The BeHEARD Addgene reagents will... lentiviral vectors from Addgene, the lab will express these candidate genes in TS iPSCs to identify whether...CTD, which arises due to mutations in the SLC6A8 gene, also known as the creatine transporter. The Association...Congratulations Mark Mellett! Siddharth Prakash: X chromosome genes involved in Turner syndrome Turner syndrome (TS)...can lead to cardiac developmental defects. Eight genes from the X chromosome are expressed at significantly...will facilitate our aims to correct X chromosome gene dosage in TS cells as a first step to reverse the...Germline mutations in one of these parts, the GNB1 gene, which codes for G𝛃1 subunits of G protein complexes...

  24. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...comparing gene activation when enhancers were active or silent, they could identify the target genes of previously...efficient. In general, epigenetic chromatin modifications are correlated with alterations in gene expression...updated Jun 24, 2020. Epigenetic modifications are an additional layer of control over gene expression that ...proteins that help package DNA, were associated with gene expression or silencing. To alter DNA modifications...oxidation and repair factors that allow for increased gene activation at the targeted site compared to other...the only CRISPR-based technology designed to alter gene expression. Fusing dCas9 to a transcriptional activator...a CRISPR-Cas9-based acetyltransferase activates genes from promoters and enhancers. Nature Biotechnology...

  25. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ...relies on the sacB gene product of B. subtilis (Hmelo et al., 2015). The sacB gene product is a sucrase... in the chromosome is to disrupt expression of a gene with an antibiotic resistance cassette. This leaves...sometimes interfering with expression of surrounding genes. However, there are ways to create scarless mutations...Red that contained one of eight bacterial toxin genes isolated from E. coli or Psuedomonas aeruginosa....toxins, and phage holins. They hypothesized that genes that had evolved specifically for toxicity would... pathogenic Proteobacteria. We placed the toxin genes under control of a rhamnose-inducible promoter, .... "Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004...

  26. Evolution of Lab Techniques

    Type
    Blog Post
    Published
    June 21, 2016, 2:30 p.m.
    ...particular gene mutation, southern blotting can be used to identify the presence of that mutated gene. In southern...the location of genes. With this knowledge, researchers were able to target specific genes for deletion ...southern blotting being used to identify a gene mutation is the genetic test used for sickle cell anemia (2)...probe (for example, has the sequence of the mutated gene the researcher is searching for), then the probe...to identify the presence of the mutated β globin gene that causes sickle cell anemia. The mutation happens...questions. Real-time PCR detects and quantifies a gene or DNA sequence of interest by recording DNA abundance...fluorescence in situ hybridization for detection of gene copy number changes." Protocols for Nucleic Acid...

  27. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ... insert the FP gene into the target gene’s native locus, but introducing large genetic changes in C. elegans...glycosyltransferase genes and other proteins related to cellular glycosylation. Get the library at Addgene CasRx ...cassette exons and disrupts genes with alternative exons. Get the library from Addgene New from the viral...library inhibits over 18,000 human genes. Find the lentiviral preps at Addgene hSyn1-SIO-stGtACR1-FusionRed...So how can you use CRISPR to introduce large FP genes into C. elegans? By using a split FP approach. Split-wrmScarlet... validated this tool by targeting six different genes whose protein products have distinct localizations... number, generally very small, and can contain different neurotransmitters. Using genetically encoded ...

  28. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...investigation into the regulation of your favorite gene. But have no fear - I’m here to introduce you to...genome – could be affected, leading to unpredicted gene expression changes (Sapozhnikov and Szyf 2021, Charlton...interact with transcriptional machinery to modify local gene expression, confounding any potential conclusions...causal relationship between DNA demethylation and gene expression (Sapozhnikov and Szyf 2021).    ... expressing blasticidin or puromycin resistance genes.  It’s that simple! Good luck and happy editing...Vogelstein B (1983) Hypomethylation distinguishes genes of some human cancers from their normal counterparts...Methylcytosine Hydroxylation, Active DNA Demethylation, Gene Transcription, and Memory Formation. Neuron 79, ...

  29. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    Published
    May 4, 2021, 1:15 p.m.
    ...., 2018). Hybridomas can lose gene expression of the mAb-encoding genes or fail to revive after cryopreservation...monoclonal antibodies that are generated in vitro using synthetic genes that are typically expressed from... integrated sequence in a stable cell line. The genes encode the heavy chain and light chain for the antibody...Figure 2: (Left) The light chain and heavy chain genes of the antibody are encoded in a plasmid. (Right...better and better at identifying and cloning the genes for antibody expression. If starting from a protein...sequences of the antibodies and then synthesize the genes that encode those amino acids (Tran et al., 2016... Antibody? animation Resources on Addgene.org Check out Addgene's antibody plasmid collection ...

  30. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    Published
    March 14, 2019, 1:54 p.m.
    ...particles. But, exo-AAVs may open new ways of improving gene delivery. They increase the efficiency of transduction...for shielding neutralizing antibodies and robust gene delivery in vivo." Biomaterials 35.26 (2014): 7598...PMC4104587. György, Bence, et al. "Rescue of hearing by gene delivery to inner-ear hair cells using exosome-associated.... "Microvesicle-associated AAV vector as a novel gene delivery system." Molecular Therapy 20.5 (2012):...PMC3345986. Meliani, Amine, et al. "Enhanced liver gene transfer and evasion of preexisting humoral immunity... "Exosome-associated AAV2 vector mediates robust gene delivery into the murine retina upon intravitreal...viral vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

  31. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    Published
    Sept. 8, 2015, 2:30 p.m.
    ...plasmids from this publication at Addgene. Multichromatic control of gene expression in Escherichia coli...optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...used to engineer new pathways with sophisticated gene expression controls. Most cell types do not respond... can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive two-component... was only about 10-fold, precluding the study of gene expression at very high or very low levels. Schmidl...researchers to better fine-tune the level of output gene expression based on the input light intensity. In...the plasmids from this publication at Addgene. A synthetic genetic edge detection program. Tabor JJ, Salis...

  32. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ...lines and are not used for general plasmid selection. Instead, reporter genes are often employed to easily... be assayed conveniently. A direct test for your gene of interest may be another method to assess transfection...conjunction with positive selection to ensure your gene gets targeted to a specific location within the ...used selection markers are listed below: Name Gene Conferring Resistance Cell Types* Mode of Action...use G418. This can be confusing since the neo/kan gene confers G418 resistance; however, like gentamicin...Read More: Plasmids 101: Antibiotic Resistance Genes Plasmids 101: Origin of Replication All Plasmids...the genetic traits of the foreign material and will subsequently pass it on to future generations. Stably...

  33. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    Published
    Jan. 21, 2020, 2:30 p.m.
    ...Imagine you’ve just discovered that your favorite gene was described in a CRISPR screen publication. You...actual data. Now you’re wondering if your favorite gene is included in other CRISPR screens and whether ...researchers to query all public CRISPR results for any gene of interest. In addition, users interested in a ... thresholds used by the authors to determine the gene hits for each screen are also captured as metadata...results are reported as the original quantitative gene-level scores provided by authors rather than sgRNA-level...available resource developed by the Biological General Repository for Interaction Datasets (BioGRID) in...Twitter at @biogrid.     Additional resources on the Addgene blog Read this blog post about genome-wide screening...

  34. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    Published
    Nov. 19, 2013, 2:37 p.m.
    ...proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed under native...sequence and a selectable gene product for isolation of successful transformants. Addgene distributes several...be interested in attaching epitope tags to their genes of interest, allowing for easy capture and detection...modifications in S. Pombe, including full and partial gene deletion, overexpression (by promoter substitution...few. Have you used these tools in your own lab? Addgene would love to hear from you, our community, about...

  35. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    Published
    May 2, 2024, 1:15 p.m.
    ...526) that (1) enables efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity...for in vivo studies; (3) is compatible with single gene KO and pooled screening approaches; (4) encodes ... optimize conditions for a screen that assessed genes that regulate antigen-specific CD8+ T cell responses...screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8+ T cell responses to ...Markson et al., 2024), developed and deposited at Addgene by Arlene Sharpe’s lab, enables in vivo screens...10.1084/jem.20230699. Additional resources on the Addgene blog X-CHIME: Context-Dependent Germline Knockout...

  36. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...REVeRT, for gene delivery using reconstitution via mRNA trans-splicing. By splitting a large gene into smaller...the introduction of extraneous genetic material to reconstitute the gene of interest, reducing the immunogenicity...hIBA1a, 466 bp) that drives high microglia-specific gene expression when incorporated in an AAV vector (Figure...doi.org/10.1101/2023.12.12.571321.  Revolutionizing Gene Therapy: The REVeRT Dual AAV Vector System By Vaibhav...AAVs, and then splicing their mRNAs, the functional gene product is reconstituted within the target cells...vector-based therapies. REVeRT supports advanced gene expression regulation, enhancing both the mechanisms...trans-splicing dual AAV vectors for (epi)genome editing and gene therapy. Nat Commun., 14(1), 6578. https://doi.org...

  37. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    Published
    March 11, 2014, 5:55 p.m.
    ...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the CRISPR/Cas9 system to precisely edit specific genes in monkeys. "Our study shows that the CRISPR/Cas9...system enables simultaneous disruption of two target genes in one step without producing off-target mutations...Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014...many human diseases are caused by genetic abnormalities, targeted genetic modification in monkeys is invaluable...invaluable for the generation of human disease models." Addgene Outreach Scientist and fellow blogger Caroline...

  38. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...variations in gene expression, they found that for approximately 40% of the identified genes, the genes were not...explanation. Next, they hypothesized that differences in gene expression could explain why some ancestry groups...groups were differentially dependent on certain genes. This didn’t prove to be correct, either. However, ...sequences for sgRNAs that target ancestry-associated genes.   What does the reference genome actually reference...cases, this may lead researchers to deprioritize genes that could be good targets for therapeutic development...cells’ germline genetic variations. They then wanted to identify cancer-associated genetic dependencies ...ancestry which will reduce the likelihood of generating genetic dependency data with an ancestry-based bias...

  39. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...on vectors where the gene of interest is inserted into the middle of the lacZ gene, thus disrupting 𝛽-...where only cells that have gained or lost a specific gene survive (ex: antibiotic resistance marker). In a... the blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which ...scientists have found that when a portion of the gene is deleted, it creates a non-functional 𝛽-galactosidase...mentioned above. In molecular biology, cloning a gene into your plasmid is often a numbers game. By using...troubleshooting Sanger sequencing results. At Addgene, we now use next-generation sequencing during our QC process....pigment. Find plasmids for blue-white screening at Addgene! Some bacterial strains contain lacZ in their genome...

  40. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...These vectors allow you to test the function of Your Gene Of Interest (YGOI) in a controlled environment under...design. You can create a new strain with a disrupted gene by transforming wild-type yeast with an integrating...modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check... More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other... is which vector will best suit your needs? At Addgene, we have a vast collection of empty backbones that...vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected cells, you will not need...
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