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We narrowed to 920 results for: Gene

Showing: 121 - 160 of 920 results

  1. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ... this gene. Should you cut it at the beginning of the coding sequence to delete the entire gene? Perhaps... in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully this post will...they have been generated (more on that later). Find Validated gRNAs to Target Mouse Genes To begin the ...chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9...decide to do a gene modification experiment in mice to the day you have your F0 generation (barring life...Haoyi, et al. "One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated ...blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use...

  2. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    Published
    Oct. 11, 2016, 2:30 p.m.
    ...knockout, activate or repress target genes in plants using the same general experimental design principles ...Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922. PLoS ONE 11:e0154027...for targeted modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR... can also be used to activate or repress target genes in plant cells, respectively. Cas9 expression is...regulate expression of longer RNAs (such as mRNAs for gene expression). Examples of commonly used RNA pol II..., activation and repression of target genes in plants and many of the aforementioned empty gRNA backbones...Agrobacterium tumefaciens as a vehicle to deliver your gene of interest into a target cell line or organism ...

  3. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ..."Future of rAAV gene therapy: Platform for RNAi, Gene Editing and Beyond." Human Gene Therapy ja (2017...required for the onset of gene expression. However, it has yet to be determined what gene expression looks like...Additional Resources on the Addgene Blog AAV: A Versatile Viral Tool for Gene Expression in Mammals A Match...transport desirable or not desirable? If diffuse gene expression is tolerated, then axonal transport is...Otherwise, using a specific system for controlling gene expression such as Cre-lox, may be sufficient to...tool to use. However, we are starting to see that gene expression really does not take much time at all... normally require second strand synthesis before gene expression begins. ScAAV avoid this rate limiting...

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

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ... to make my first CRISPR/Cas9 gene edit. I wanted to inactivate the gene BRAF (a kinase implicated in ...Targeting different regions of the gene is also useful in case the gene has a weird mutation that you didn...each step of performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic...Schematic overview of CRISPR/Cas9 gene editing process. See below for details. The entire process from start...what level of expression was the most efficient for gene editing. A virus encoding GFP plus a gRNA targeting...GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9 should appear green and ...multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA viruses, but you can ...

  5. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...especially when fused to effector proteins for gene activation or gene editing. The more recently identified Cas12f...SEND for gene transfer with pseudotyped viral particles, expanding options for CRISPR or other gene delivery...editing systems improve gene editing performance at therapeutically relevant gene targets and expand the...version (named CasMINI) had the highest activity for gene activation  and base editing, whereas a different..., and fusogen in a modular system for delivering gene editing tools into cells. Image adapted from Segel...editing to correct mutations in a variety of target genes and cell types. Their Nature Biotechnology paper...tyrosinemia, where a pathogenic insertion within the FAH gene was precisely corrected, restoring expression in...

  6. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ... transcription of a target gene, it therefore determines the timing of gene expression and largely defines...synthesis and allowing reporter gene transcription to take place. Addgene also has popular light-regulated...the promoter and ceases repression of the target gene. A simplified lac inducible system is found in many...shock-inducible Hsp70 or Hsp90-derived promoters, in which a gene of choice is only expressed following exposure to...inducible promoters Light is another way to activate gene expression, and two-component systems used in synthetic...promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, preventing...experimental system. Leakiness If you’re expressing a gene that may be toxic, it’s important that your inducible...

  7. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    Published
    June 4, 2015, 1:03 p.m.
    ...the plasmid containing the lethal gene. An example is the ccdB gene system commonly used in Gateway® Cloning...are rescued by genes supplied on the transformed plasmid. Such plasmids may contain genes that allow for...auxotrophic cell lines), or genes which rescue a lethal phenotype. Resources on the Addgene Blog Read Our Post...well-characterized bacterial lac operon contains a gene called lacZ that encodes for the enzyme β-galactosidase...discovered that deleting a section from the lacZ gene (a mutation called lacZΔM15) creates a non-functional...other methods. Positive selection vectors encode a gene which, when expressed, is lethal to the cell. Cloning...fragments are inserted into an MCS in the center of this gene, disrupting the lethality. This is similar to α-...

  8. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ...number of genes involved in cardiovascular and metabolic diseases. We are knocking out genes that have...and find several mutations in the same gene or in multiple genes, all of which seem to contribute to a ...versions of 15 different disease-associated genes. Addgene spoke to Musunuru about how the new kit works, ...scenarios, you can either knock out a gene or knock in a mutation. Addgene: How do you use this kit to target... paper, for example, we generated hepatocytes to examine the effect of a gene on cholesterol metabolism...They do that by studying patients to uncover new gene variants associated with conditions of interest,...be advantageous if you are trying to knock out a gene. One can also exploit homology-directed repair by...

  9. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...used in the lab, the transposase gene is provided in trans so that a gene of interest can be inserted between... 2021).  Gene transfer Transposons are an alternative to viral vector delivery of transgenes, like for...response and are also easier and cheaper to generate. Gene disruption due to integration can occur for...several different applications (mutagenesis screens, transgenic animals, gene transfer). The discovery of ...bacteria, transposons can move antibiotic resistance genes between plasmids or from plasmids to the bacteria... genome. In humans, if a transposon jumps into a gene, it could be mutagenic and result in human disease... and mechanism of replication. They contain two genes: gag and pol. The pol polyprotein encodes the reverse...

  10. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...to investigate the roles of specific genes or to knock down genes with potentially harmful mutations in...target gene but have a lot of off-target effects. However, this is a common issue with many gene silencing...this could be applied to. When it comes to altering gene expression, CRISPR technologies have gradually taken...) is, as the name might suggest, an RNA-mediated gene silencing mechanism. RNAi was a landmark discovery...focusing on shRNA, one of the most popular choices for gene silencing. RNA interference RNAi is a useful tool... in therapeutic settings. Silencing of a target gene is achieved through the delivery of a double-stranded... these tools. Next time you need to knock down a gene, maybe you’ll consider using RNAi! References and...

  11. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...specific genes are signified with a minus sign (-) as is typically standard; just having the gene listed... of the more common genetic changes found in E. coli strains. Table 1: Common gene mutations found in ...carefully selected the optimal promoter for your gene of interest, painstakingly cloned into the perfect...backbone, made sure to add the right tags to your gene, and may have even put a fluorescent protein downstream...in E.coli strains Gene(s) Description Functional Consequence dam DNA adenine methylase mutation (GATC...methylation sensitive. dnaJ Mutation in a chaperonin gene Increases the stability of certain expressed proteins...encodes proteins needed for bacterial conjugation. Genes listed on F´ are wild-type unless indicated otherwise...

  12. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    Published
    May 19, 2016, 2:30 p.m.
    ...aberrant activation of nearby genes. These safer vectors have become standard in gene therapy. Researchers have...condensed to create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very...into the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating (SIN... LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment of a molecular... Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources on the Addgene Blog ...applications, and they’ve also had success in clinical gene therapy applications. Read on to learn more about...researchers are interested in developing these vectors for gene therapy. In 2003, VIRxSYS began the first clinical...

  13. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...MSCV (Murine Stem Cell Virus) sequences. Packaging genes (viral Gag-Pol): Gag is a structural precursor protein... protein, and Pol is a polymerase. Envelope gene (may be pseudotyped to alter infectivity): The use of...both lentiviruses and γ-retroviruses use the same gene products for packaging (Gag, Pol, and Env), the ...and/or Env, eliminating the need to deliver these genes in trans by specific plasmids. This method, therefore...standard γ-retroviruses use the Gag, Pol, and Env genes for packaging; however, they are different viruses...replication incompetent? Yes, as with lentiviruses, the genes required for γ-retroviral packaging and transduction...U3 region of the 3’ LTR and are primarily used in gene therapy research to reduce promoter interference...

  14. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...multiple gRNAs targeting a single gene can increase the likelihood of gene knockout, or multiple gRNAs may...genome editing has been used to study the effect of gene knockout in vivo and in vitro, as well as to insert...observe a phenotype, the modification of multiple genes is often necessary. Guide RNAs (gRNAs) are commonly... al constructed polycistronic glycine tRNA-gRNA genes (PTGs) in the forms tRNA-gRNA and tRNA-gRNA-tRNA...constructed PTGs containing 8 gRNAs targeting 4 MAPK genes, again using rice protoplasts. Each pair of gRNAs...may be used to target genes with overlapping activity. Multiple gRNAs can also facilitate deletions of...viruses have illustrated the power of polycistronic gene organization, and its translation to CRISPR/Cas9...

  15. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ... or “LSL” cassette) upstream of a gene of interest will prevent gene expression in the absence of Cre....Cre-dependent gene knockout – conversely, putting the loxP sites on either side of a gene (called “floxing... loxP”), will permit gene expression until Cre is present, at which time the gene will be disrupted or...most common uses are listed below. Cre-dependent gene expression – placing a series of transcriptional... presence of Cre, the stop codon is excised, and gene expression proceeds. One popular lox-stop-lox plasmid...switch regions evidenced through Cre–loxP–mediated gene targeting. Gu, H., Zou, Y.R., and Rajewsky, K. 1993...reciprocal crossover event. This event can have three general outcomes based on the location and orientation ...

  16. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    Published
    March 24, 2016, 2:30 p.m.
    ... (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 ...

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

    Type
    Blog Post
    Published
    April 7, 2015, 4:21 p.m.
    ... 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...

  18. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...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 ...

  19. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...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...

  20. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...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...

  21. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

    Type
    Blog Post
    Published
    Dec. 12, 2013, 3:35 p.m.
    ...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...

  22. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 3:42 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    April 23, 2018, 2 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ... 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...

  28. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...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...

  29. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Sept. 9, 2016, 2:43 p.m.
    ...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...

  31. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    Published
    March 21, 2023, 1:15 p.m.
    ...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. ...

  32. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...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...

  33. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    Published
    Nov. 15, 2016, 3:30 p.m.
    ...explains how gene drives work and what work he's planning on doing with CRISPR-based gene drives focusing...developing new CRISPR tools, and the potential uses of gene drives. Listen here! Podcast breakdown (Time : ...- 31:25 : Nature's defenses against gene drives. 31:26 - 33:08 : Plans for future work at the Wyss Institute... gene drives...became a member of the Addgene advisory board where he will help shape Addgene's future. Listen to this...member of the Addgene Scientific Advisory Board with a particular interest in generating Cas9 based tools...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...

  34. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    Published
    Oct. 31, 2019, 1:06 p.m.
    ...lifecycle. Halloween genes The Halloween genes are a set of cytochrome P450 genes discovered in Drosophila...Mutations in these genes are lethal, associated with defective exoskeleton formations. These genes were appropriately...1983): 813-816. Gilbert, Lawrence I. "Halloween genes encode P450 enzymes that mediate steroid hormone...We’ve had teams dressed up as plasmids, Blugene, and the Addgene ping pong table. This year, in addition...Additional resources on the Addgene blog Read about company culture at Addgene See some of our previous ...costumes, and spooky science stories. Yes, please! At Addgene, we love finding ways to incorporate science themes...Halloween contest, we’ve also carved pumpkins (yes, Blugene made an appearance there too) and shared fascinatingly...

  35. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...mixture of XFP genes. Stochastic multicolor labeling is possible when each of the XFP genes is introduced...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced...result in a combinatorial expression of these three genes with different copy numbers, producing dozens of...modern neuroscience, plasmid or viral vector-mediated gene delivery is more common than conventional transgenic...transgenic strategies. In such vector-based gene delivery systems, the stochastic expression of XFPs is possible... Plasmid concentrations and AAV titer - The XFP genes have to be introduced at ~2 copies per cell per ...plasmids (Addgene #104102 - #104105) using in utero electroporation or with AAVs (prepared with Addgene #104110...

  36. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...varies across targets, they tested two genes in mice. For both genes, they saw indel formation and phenotypic...in vitro and germline genome editing, but in vivo gene editing approaches have been limited by problems...viral vectors (AAV) are commonly used for in vivo gene delivery due to their low immunogenicity and range...al. successfully used CjCas9 with AAV to target genes in mouse muscle and eye tissue. Neisseria meningitidis...cell is transduced by both AAVs, the full length gene and/or protein is reconstituted. One way to reconstitute...split intein spCas9-AAV toolbox that retains the gene-targeting capabilities of full-length SpCas9. This... of plasmids includes AAV-Cas9C-VPR for targeted gene activation. Split inteins have also been used to...

  37. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...knockout of a target gene and expression of a separate gene. In these mouse lines, gene knockout can be induced... type specific) control of DNA recombination and gene expression can be advanced by the coordinated use...recognizes FRT sites and initiates site directed gene recombination between those sites. FRT sites and...sequence of interest can lead to poor expression of genes within that sequence and should therefore be prevented...recombination of the loxP sites and thereby to the gene knockout. In this experiment it is possible to compare...morphology and synaptic connectivity before and after the gene knockout in the same mouse.   Scientists always...researcher exquisite control over expression of multiple genes in specific cells at specific times. Many thanks...

  38. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...CRISPR reagents and other specific gene sequences available from Addgene. Dr. McMurray hopes that by demonstrating...lab has generated mice wherein it's possible to delete the functional portion of the Nfix gene using Cre...phosphomannomutase (PMM) activity due to mutations in the PMM2 gene. Videira’s group recently reviewed and reported ...interactions and how these interactions regulation gene expression. Recent studies have shown that the conditions... Smith Syndrome are due to mutations in the NFIX gene. These diseases are characterized by overgrowth ...studying the function of the mouse Nfia, Nfib and Nfix genes in brain development since 2000 and have shown several...NFIX function. They will then mutagenize the NFIX gene and test whether variants of unknown significance...

  39. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...understanding of B. subtilis gene functions and interactions. The single-gene deletion library is derived...studying the phenotype of an essential gene, the CRISPRi Essential gene Knockdown library was constructed ...Bacillus subtilis libraries with Addgene – the B. subtilis Single Gene Deletion Library—Kanamycin and the...compatible cloning sites for easy cloning of your genes of interest. In order to improve the study of specific...cassette. However, it was unknown whether variations in gene expression from these polycistronic cassettes could... stoichiometry impacted reprogramming and global gene expression patterns. Polycistronic cassettes that... the CRISPRi Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many...

  40. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...found in the same gene. If we only have small, directed editing abilities, any gene editing treatment ...functional copy of the entire gene, allowing a one-size-fits-all approach to gene replacement therapy for most...insertions, leaving a several kilobase-sized gap in the gene editing field. The AbuGoot lab has now developed...to work. (It was also found to be independent of gene repair.) This lowers the chance of the insert being...activity. They now had a system that could integrate genes up to 36 kb in length, with an integration rate ...system.     Fig. 2: Schematic of programmable gene insertion with PASTE. The PASTEsystem involves insertion...primary cells, and can be easily programmed for new genes. In comparison with other large insert techniques...
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