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We narrowed to 1,144 results for: Gene

Showing: 261 - 280 of 1144 results

  1. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

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    Blog Post
    ... (NHEJ) pathway disrupt target gene sequences (thus inactivating genes), while breaks repaired by homology...error-prone repair of the gene would be disastrous (i.e. if breaking the gene is worse than leaving the...(in our hands) this technique supports gene disruption or gene replacement frequencies that are an order...repair (HDR) pathways can modify the sequence of a gene (thus altering its function). HDR is crucial for...consistently supported higher frequencies (up to 60%!) of gene editing than single or double stranded donor DNA...editing, but it could be very useful to tackle genetic diseases in which sequence replacement holds some...PubMed Central PMCID: PMC4032847. Resources on the Addgene Blog Brush Up on Your HDR Basics Learn How to ...

  2. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ... delete/overexpress endogenous genes or to introduce heterologous genes, but the trend of research has...tinkering with regulatory networks and multiplex gene control. However, these approaches are often met...mimic natural transcriptional programs on multiple genes. Modifying the CRISPRi system for multidirectional...programs to differentially alter the levels of five genes that encode biosynthetic enzymes for violacein, ...recruit various transcriptional regulators to alter gene expression at the targeted site. Multiple scaffold...simultaneous, multidirectional and tunable control of gene expression. This strategy can be applied to effectively...RNA-guided platform for sequence-specific control of gene expression. Cell, 152(5), 1173-1183. (2013). PubMed...

  3. Deep Dive: qPCR

    Type
    Blog Post
    ...profiles (Dwight, 2011).  Reference Genes  Choosing a set of reference genes deserves more attention than it...reference genes, plural. Your data will be much improved if you move to using multiple reference genes whenever...reference gene being a common choice - the frequently used GAPDH has noted issues as a reference gene (Higgins... Validation of Common Housekeeping Genes as Reference for qPCR Gene Expression Analysis During iPS Reprogramming... it usually receives. Different reference genes have shown to have as much as sixfold variation in expression...), so you’ll want to ensure that your reference genes are suitable for your cell lines and/or tissue types..., and it's possible to multiplex your reference genes to save on plate space. For many cell lines and ...

  4. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...easily modified to tag hundreds of genes, generating powerful genetic screening libraries. Additional protein... to screen for the genetically modified strains that contain your newly tagged gene. Most insertions can... to generate high-throughput targeting vectors. These vectors can be used to tag endogenous genes and ...-Throughput CRISPR/Cas9 Gene Modification in Caenorhabditis elegans.” Genetics. 202(4) (2016):1277-88....protein fusions is to replace the genomic copy of your gene of interest with a fluorescent protein fusion. CRISPR...allowed scientists to tag and light up endogenous genes of interest to better understand normal protein ...easily create C-terminal tag fusions of endogenous genes in human cells. To use CRISPaint, users need 3 vectors...

  5. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...that of adjacent cellular genes. Q7: Which genes are deleted or modified to result in replication deficiency...including the gene of interest, and the plasmid coding for an envelope. Three generations of lentiviral...minimal number of HIV genes required for virus production (3 or 4). Third-generation vectors contain gag...gag, pol, and rev. Second-generation contains those three genes plus tat. The envelope plasmid provides...are useful and efficient tools to introduce your gene of interest into cells. Unlike gamma-retroviruses...lentiviruses and gamma-retroviruses use the same genes for packaging (that's gag, pol, and env), the isoforms...infect dividing cells. When choosing the appropriate gene delivery system, the specific target cell or tissue...

  6. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    ...forward engineer a system of 10 genes. You can now forward engineer a 10-gene system to do something. We know...collection of genetic parts for precisely forward-engineering the central dogma across many genes simultaneously..., off-the-shelf genetic elements that allow you to begin to imagine designing 10-gene systems with quantitative...used over and over again in combination with novel genes to get very precise expression levels. The BIOFAB...otherwise invisible molecules into regulation of gene expression including growth or fluorescence. There...our testing of these parts that the chance your genes will be expressed at the level you want is 93 percent... of biological systems today who are building 10-gene systems from scratch. (They may be building genomes...

  7. Zinc Finger Consortium Reagents

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    Collection
    ...Arrays for Zebrafish Genes - Browse pre-constructed zinc fingers targeting specific genes Oligomerized Pool...Keith Joung and Daniel Voytas have deposited at Addgene various reagents for engineering and expressing...

  8. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    Blog Post
    ...Cardiac Gene Transfer Superior to AAV1, AAV6, AAV7, and AAV8 in the Mouse and Rat. Human Gene Therapy,...toolkit for human gene therapy. Molecular Therapy: The Journal of the American Society of Gene Therapy, 14(...AAV1 AAV1 was the first viral vector approved for gene therapy (Issa et al., 2023). It may be a good choice...exposure or rAAV treatment, which can pose problems for gene therapy applications. Cross-reactivity of AAV serotypes...adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous. Science Translational... Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), 785. https:/...Ayasoufi, K., & Barry, M. A. (2019). Comparison of Gene Delivery to the Kidney by Adenovirus, Adeno-Associated...

  9. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...physiological gene expression regimes that cannot be easily achieved with classical gene overexpression...contain changes in expression of TRIM21-associated genes, indicating that TRIM21 overexpression does not ...variety of protein modules to repress or activate gene expression, sometimes incorporating chemically inducible... 28916764. Plasmids for independent control of gene expression in yeast Article contributed by Tyler...that can independently control expression of your genes of interest. These transcription factors consist... appropriate promoter sequence and activation of gene expression. In addition to plasmids encoding these...and ZIF268. To demonstrate independent control of gene expression, Aranda-Díaz et al transformed yeast ...

  10. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...their test genes, the researchers at the BIOFAB showed that expression of many different genes was predictable...translational repressor to down-regulate a gene of interest. You can generate PUF proteins with a wide variety ...restriction enzymes and standardized primers to clone your gene of interest into both sequencing (pInitial) and ...abundance (meaning they are unlikely to be found in your gene of interest) and prevent incorporation of extensive...kit contain a variety of promoters to facilitate gene expression in bacteria, yeast, mammalian cells, ...hope this kit will allow you to quickly study your gene of interest in a variety of model systems, thereby...peptides) downstream of which you can clone your gene of interest. Translation of the standardized leader...

  11. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ...viral (AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal...organs to the CNS, are also an important target for gene delivery, especially for the study of pain. While...all possible combinations into a region of the cap gene. In this special CREATE cap plasmid, loxP sites ...primers to determine which version(s) of the capsid gene successfully infected the cell type of interest....translational applications (mechanistic studies and gene therapy treatments for neurological diseases). The...capsids can serve as powerful cell-type-restricted gene vectors for neurons, no transgenic mouse required...et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems...

  12. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ... what part of a gene enhancers like to interact with, and how many promoters or genes each enhancer is...plot line in gene regulation. When chromatin conformation data is compared with changes in gene expression...dynamic role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated with CAPTURE...changes in gene expression and determine who’s a key driver of these changes. By generating a more vivid...updated Jul 2, 2020. Promoters may be the star of gene regulation, but enhancers and chromatin looping ... bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers are located ...located thousands of base pairs away from the gene they regulate, but are brought in proximity by the looping...

  13. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ... one bacterium sharing genes with another, an accidental transmission of genes from one bacterium to another...‘horizontal gene transfer’. Resistance can also arise from random mutations in the genetic sequence that...Additional Resources on the Addgene Blog Learn How Antibiotic Resistance Genes Are Used in the Lab Probe...consuming to treat. Learn How Antibiotic Resistance Genes Are Used in the Lab How does antibiotic resistance...bacterial cells), or a bacterium picking up resistance genes from it’s environment (from DNA left behind after...develop, either through random mutation or horizontal gene transfer, means to defend themselves from these ...while the few that happen to possess resistance genes survive and multiply. Factors contributing to the...

  14. CRISPR Antimicrobials

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    Blog Post
    ...resistance gene, allowing resistant bacteria to multiply, spreading their resistance genes throughout ...them to multiply and propagate their resistance genes. Our society is hoping to combat antibiotic resistance...bacterial species, or even an antibiotic resistance gene. Importantly, native, non-pathogenic bacteria remain.... targeted beta-lactam and quinolone resistance genes in E. coli using plasmid and phagemid delivery systems...population to the antibiotic, despite this resistance gene being encoded by a high-copy plasmid. In the case...CRISPR machinery targeting 1) the given resistance gene and 2) a second lytic phage. Bacteria that receive...work targeted two distinct beta-lactam resistance genes, again with very high specificity. This double phage...

  15. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ... containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral ...Talking 'bout my (lentiviral) generation The first step in delivering genes via lentivirus is deciding what... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges...split the viral genome by removing the viral Rev gene from the packaging plasmid and incorporating it ... the structural elements, envelope, and transfer gene are transfected into a packaging cell line, usually...) while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how...sufficient titer; this is often the case with large genes. This is due to inefficient packaging of large inserts...

  16. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    ...screens using CRISPR technology, gRNAs to knockout genes – are essential screening components, the model ...Sometimes, you might have a few positive control genes on which to test the validity of your biological... a new assay or both, and you have no idea what genes are involved. Or you don’t know how well your assay...definition, the kinome is a well-annotated collection of genes that can provide a useful starting point for developing...screens can be performed with either 4 or 8 gRNAs per gene. Further, the pools are available in two vector ...individual plasmids will also be useful for imaging or gene expression assays necessitating the screening of...blogger, member of the Addgene Advisory Board, and Associate Director of the Genetic Perturbation Platform...

  17. Synthetic Biology - Cloning and Genomic Tools

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    Collection
    ...Metabolism Networks & Gene Regulation Sensing & Signaling Strains Browse Addgene's collection of synthetic...available from this depositor's lab. Plasmid Description Gene/Insert Vector Type PI Publication Back to Top Shuttle...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...

  18. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...was previously a large barrier for gene therapy. In fact, AAV gene therapies have already been approved...Transient Gene Expression High Immunogenecity AAV Remains Episomal (Little Risk of Mutagenesis) Very...of Insertional Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity (...Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy with...Type of Virus Pros Cons Retrovirus Stable Gene Expression Broad Tropism Only Infects Dividing...Immunogenicity Infects Non-Dividing Cells Transient Gene Expression Low Tropism for Mouse Cells and Hematopoietic... lentiviral vectors through pseudotyping.” Curr. Gene. Ther. 5 (2005) 387-398. PubMed PMID: 16101513. ...

  19. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...of homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids used...host mutations by including a copy of a functional gene which complements the host’s auxotrophy. When grown...have incorporated the plasmid carrying the required gene.  The table below lists some of the most commonly...growth-based positive selection for the loss of the marker gene) For use in: Can this be used in other yeast species...yeast phenotypes. This marker should not be used in gene disruption experiments. MET15 L-methionine and...FOA in fission yeast induces mutation in the ura5+ gene in addition to ura4+. leu1+ L-leucine   S. pombe...issues. One method to reduce the amount of marker gene expression is to use a partially defective promoter...

  20. Retrovirus Plasmids

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    Collection
    ...your gene fused to eGFP expression Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression...transfer plasmids. Want a retroviral plasmid with your gene of interest? Search our database for retroviral ...Puro IRES GFP empty plasmid) MSCV For cloning and gene expression; select with puromycin or screen for ...'s genome, which can lead to oncogenesis (termed insertional mutagenesis). Read our γ-Retrovirus Guide...Learn about retroviral plasmids at Addgene, including popular packaging, envelope, and transfer plasmids...virus uses the reverse transcriptase in tow to generate a DNA provirus. This provirus then gets integrated...to ∼8 kb), and infection results in long-term transgene expression. Some disadvantages of γ-retroviruses...
Showing: 261 - 280 of 1144 results