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We narrowed to 920 results for: Gene

Showing: 241 - 280 of 920 results

  1. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...permanently knock out or knock-in a gene? Do you want to activate or repress gene expression? Are you trying to...reporter gene (e.g. GFP) to identify and enrich positive cells or a selection marker to generate stable ...conjunction with activator or repressor domains to control gene expression. Base editors can help you make precise... separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells... for each, and potential delivery methods.   General considerations for planning your CRISPR experiment...Wildtype Cas9 or Cas9 nickase are appropriate for generating knockouts or knock-ins, or introducing mutations...PCR followed by gel electrophoresis and/or next-generation sequencing more appropriate? If you already...

  2. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...effectively targets your gene/region of interest is a key step for CRISPR/Cas9 gene editing. Dr. Masahito...sequence or gene of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers...vector and gene primer for SLIC cloning. Amplify the pCoofy vector and the sequence of gene of interest... can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters, or other...Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab has...approach to genetic engineering which drastically reduces the time and effort needed to insert new genes into...successfully create GFP fusion proteins with a variety of genes across the genome. Although this tool was published...

  3. Plasmids 101: Origin of Replication

    Type
    Blog Post
    Published
    Nov. 11, 2020, 2:23 p.m.
    ...Now that we know all about antibiotic resistance genes, let’s consider another basic element of any plasmid...Do you want to make a lot of your plasmid? Is the gene toxic in high amounts? It is always good to keep...below for additional considerations. *Requires pir gene for replication (reference). **F1 is a phage-derived...copies of plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain...survive within cells. The replicons of plasmids are generally different from the those used to replicate the... to make additional copies. ori sequences are generally high in As and Ts. Why, you ask? Well, A-T base...few copies depending on how they are regulated. Generally, control of replication is referred to as "relaxed...

  4. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    Published
    Dec. 21, 2017, 2:06 p.m.
    ... extensive heterogeneity in the feature sequences due to genetic drift and the use of genes from different...raw plasmid sequence and display frequently used genes and control elements. Development of this tool required...sequence feature that may be used to make fusion genes, detection needs to occur even if one or two codons...of the feature. With the increasing popularity of gene synthesis, many researchers now use codon-optimized... who uses SnapGene or the free SnapGene Viewer companion product. If you don’t use SnapGene yet, you can...Resources on the Addgene Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the...sequencing at Addgene Additional Resources on Addgene.org Example plasmid analysis powered by SnapGene Browse...

  5. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...plasmid to ensure our assembly result is accurate The gene/insert or any major feature highlighted by the deposited...sequenced by Addgene’s QC team. Email help@addgene.org to get started! Addgene’s QC process Addgene prioritizes...deposited sequence(s) in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature” library...Quality Control Now at Addgene Bioinformatics at Addgene Resources on Addgene.org Educational Resources... Figure 2. Addgene’s basic QC process. Created with BioRender.com. Once Addgene receives deposited...receiving the replacement plasmid at Addgene. QC is a team effort Addgene’s QC team is full of dedicated scientists... Resources Additional resources on the Addgene blog Addgene's Tips for Plasmid Quality Control Plasmids...

  6. Addgene's Website Update

    Type
    Blog Post
    Published
    April 8, 2024, 2:15 p.m.
    ...do scroll down? You’ll find Blugene, as well as information about Addgene, our mission, links to our deposit...improving the tools you use to share and access them. Addgene’s product, development, and content teams have made... made some changes to the look and feel of addgene.org. Some you’ll notice right away, while others may...resources, tools, collections in our catalog, and the Addgene Help Center. A big part of this redesign was pulling... place, so you can quickly navigate around the Addgene website without scrolling up and down the page....skim through. Users with disabilities will find Addgene’s website easier to navigate, thanks to design choices...— check out all the new features and designs addgene.org! And when you’re finished, feel free to take ...

  7. Summer Fun at Addgene!

    Type
    Blog Post
    Published
    May 12, 2022, 1:15 p.m.
    ...means summer is right around the corner. Here at Addgene, we’re always inspired by the new seasons, so we've...complex questions, and giving tips for consistently generating solid data. If you’re ready for the next steps...We’re very excited to be launching the official Addgene TikTok at the end of this month! We’ll be sharing...lab, along with some behind-the-scenes peaks of Addgene’s people, culture, and workplace. Rumor has it that...

  8. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...targeting EvolvR to five dispersed regions of the target gene of spectinomycin, rpsE. These results not only uncovered...directed mutagenesis to generate specific mutations in your plasmids Use REPLACER Mutagenesis to mutate...and forward genetics harnesses this at a large scale by screening diverse libraries of genetic variants....Berkeley, provides a new approach for generating novel genetic variants in bacteria. Read on to learn about...EvolvR’s on-target mutagenesis rate was 2.5 x 10-6 mutations per nucleotide per generation vs. 10 x 10-10 ...additional mutations to PolI3M generated PolI5M which increased EvolvR’s mutagenesis rate to ~10-3 mutations ...than one genomic site for mutagenesis, a feature which they used to generate bacteria resistant to both...

  9. Lab Automation at Addgene

    Type
    Blog Post
    Published
    March 22, 2018, 1:28 p.m.
    ...mail due to laws concerning genetically modified organisms. Since Addgene is very proud to help scientists...times easier. We’ve certainly felt this way at Addgene so we’ve been putting some work into finding robots... many more robots than you ever knew existed! Addgene recently started automating a few protocols to ... hour. Automating vial fiIling The majority of Addgene plasmids are shipped as agar stabs in glass vials... easily programmed for a variety of lab needs. Addgene uses it to fill as many vials as it can in the ...Label your tubes with a label maker. Label makers generate far more legible labels than your hand and help...avoid sore fingers. There are other process that Addgene would love to move forward and automate. For instance...

  10. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral helper sequences...viral vectors are tested in vitro and in vivo for gene expression and/or function. These data are sometimes...copies of the lentiviral Rev responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit...vector genome titers by droplet digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://doi.org...Titer by a Novel Droplet Digital PCR Method. Human Gene Therapy Methods, 29(2), 96–103. https://doi.org/...Production AAV distributed by Addgene has been produced either in-house by Addgene scientists or through collaboration...either performed by Addgene or by the University of Pennsylvania Vector Core. In general, titering is performed...

  11. Countdown to Halloween @Addgene

    Type
    Blog Post
    Published
    Oct. 29, 2014, 5:26 p.m.
    ...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...email system.  Our 2013 winners dressed up as Addgene's favorite lunch... a BonMe noodle meal, crammed...interested in finding out who wins this year’s Addgene costume contest, like us on Facebook and checkout...  addgene...

  12. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...these expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...matters is the number of hits to the same sequence (gene). In a clean sample, there are typically fewer than... However, a large number of hits to one sequence/gene - we set our threshold at >100 hits - immediately...example, AAV-68544 contained 220 hits to the CHRM4 gene. When contamination is suspected, we proceed to...Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org/10.1089/hum...purity. But Addgene is also conducting more rigorous quality control, including next generation sequencing... for transgene expression. Once packaged in AAV, this pre-recombined DNA could lead to transgene expression...

  13. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...-invasive systemic gene delivery in non-human primate (NHP) brains.  Systemic gene delivery capsids previously...Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific ...Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain....plasmid here! Chen, X., et al. (2023). Functional gene delivery to and across brain vasculature of systemic...past decade, primary cells remain difficult to genetically edit due to toxicity from electroporation or ... under CC-BY license.   The Miyawaki lab generated a series of StayGold mutants and identified a ...mentioning that Kiryl Piatkevich and colleagues generated yet another monomeric StayGold variant (Piatkevich...

  14. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...her work has always been focused on gene transfer, from transgenes to plants, chloroplast to the nucleus...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have ...The MoClo Plant Kit makes it really easy to clone genes for expression in plant cells. Sylvester and I chose... Now there are many, so if you want to make a 10-gene construct, you don’t have to worry about repeating...constructs to allow people to, for example, knock out a gene with simultaneous complementation, with a knockout...Marillonnet share with the scientific community via Addgene, the struggles of plant scientists and how they...Everybody's favorite parts for plant synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning ...

  15. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    Published
    May 15, 2014, 3:33 p.m.
    ...from that of the gene of interest, or expressed from the same transcript as the gene of interest, but ...visualize when and where the gene is expressed. Click here to view Addgene’s collection of empty backbones... a wide number of functions, including: tagging genes for elucidating their expression or localization...promoter of interest can be used to effectively monitor gene expression from that promoter in a given cell type...more about optogenetics at OpenOptogenetics and find optogeneic actuators and sensors at Addgene. Cell marking...and others. Review Addgene's collection of fluorescent biosensors here. Optogenetics: Scientists can use...implantation and generation of transgenic mice. Purification: GFP can be used as a general epitope tag for...

  16. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...dealing with a defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a ...Adenine base editors (ABE) What if you have another gene Y, which has a defective allele Ym and you need ...first created a defective chloramphenicol resistance gene (CamR) by introducing a point mutation (H193Y). ...transformed it into E. coli harboring the defective CamR gene. Screening for surviving colonies and subsequent...system. We use a single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables... slim chance of generating indels with base editors, keep in mind that the generation of indel sometimes...biology Resources on Addgene.org Find all CRISPR plasmids and resources at Addgene Find CRISPR base editors...

  17. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...ampicillin resistance gene, you’ll find the full name of the gene, the name of its gene product, and a description... plasmid features such as antibiotic resistance genes, selectable markers, promoters, origins, tags, and... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...scientists) for suggesting these changes, and the SnapGene and Addgene software development teams for working tirelessly...example below. Our old map is on the left while the SnapGene powered map is on the right (click here to see...

  18. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    Published
    April 4, 2023, 1 p.m.
    ...for its powerful promoter of a gene important for methanol usage. This gene promoter, AOX1, was co-opted...experimental model. In fact, approximately 25% of yeast genes have a human counterpart, enabling study of human-relevant...Saccharomyces cerevisiae to human: The important gene co‑expression modules. Biomedical Reports 7.2 (2017...fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first eukaryote (S. cerevisiae... new at all, but it’s not as well-known in the general research community. Phillips Petroleum Company...release the K. phaffii expression system to the general research community in the 1990s when they abandoned...the strain tractable for peroxisomal studies on biogenesis, degradation, and proliferation. Genome manipulation...

  19. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    Published
    March 30, 2023, 1:15 p.m.
    ... for gene expression, turning transcription on and off by binding proximal to their target genes. Since...Since gene expression patterns determine everything from stem cell differentiation fate to tumor suppression...from the library to further investigate specific genes and TF isoforms. Due to the size of the collection...use to generate the relevant disease model. Accurate modeling of a disease depends on both genetic and expression... not yet available, just ask an Addgene scientist at help@addgene.org! References and Resources  References...scientific community. Recently, the Zhang lab generated a TF library (aka the MORF Collection) and an ...isoforms transcribed from these sites. The Zhang lab generated a comprehensive library of each unique TF open...

  20. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ... popular tools for genetic screens, enabling genome-wide interrogation of the genes required for cellular...that constitutively express Cas9 and screen for genes that affect Th2 differentiation and activation, ...inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the researchers explore many...-LPs and accelerate the discovery of additional genes and small-molecule probes to aid in the understanding...transduction, now enabling forward genetic screens in cells with few genetic tools. Find the mouse CRISPR ...plasmids at Addgene. Liu et al., Nature Communications, 2019. PubMed PMID: 31451697. Genetically encoded ...responsible for a phenotype can be identified by next-generation sequencing of the genomic DNA. However, some ...

  21. Addgene Stands with the Scientific Community

    Type
    Blog Post
    Published
    April 23, 2025, 1:30 p.m.
    ...utilizes grants to support our own initiatives, Addgene is deeply concerned about these developments. These... We Are Here to Help Despite these challenges, Addgene remains steadfast in our mission to accelerate ...vital during this period of constrained funding. Addgene is uniquely positioned to help research institutions...Universities and research institutions can turn to Addgene as a trusted partner for archiving their biological...scientific progress depends on our collective response. Addgene stands ready to collaborate with partners across...work and what support would be most valuable from Addgene during this time. Your input will directly shape...scientific community. Let us know by emailing help@addgene.org. We look forward to hearing from you!...

  22. Addgene: The First Twenty Years

    Type
    Blog Post
    Published
    Dec. 19, 2024, 2:15 p.m.
    ...allow people to add a gene to a repository others could request from — Addgene!  Addgene was established as...information across the globe.  “Read the Addgene guide: https://www.addgene.org/guides/crispr/” — SirHamhands, ... which continue to grow today. “I generally refer people to Addgene for the basics of molecular biology...celebrating Addgene's 10 year anniversary.   Expanding our work In March 2015, Addgene reached the...a resource like Addgene to all get the same virus.” — Grad student visiting Addgene’s booth at the Society...Addgene is ending 2024 with big plans for our future. And while we’ve been excited to share them with...thoughts from the community that makes us… us. “The Addgene not-for-profit is an amazing service to biology...

  23. Introducing...Addgene Quick Tips!

    Type
    Blog Post
    Published
    Sept. 19, 2024, 3 p.m.
    ...perhaps are a little limited on time — check out Addgene’s new Quick Tip series on YouTube! In this series...

  24. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ... page of the Addgene Data Hub.   Antibody data reports can be searched and sorted by gene/target, name... on addgene.org Addgene's Antibody Collection Addgene's AAV Data Hub More resources on the Addgene blog... request for your experiment!  Addgene’s Antibody Data Hub Addgene’s Antibody Data Hub can help with ...open-access Data Hub, researchers interested in the Addgene antibody collection can look at user-deposited ...experimental conditions in which the data was generated, you can scroll down the page to the materials...each data submission we receive is checked by Addgene's curation team for accuracy and completeness. The... citation. If you’re considering requesting an Addgene antibody, the Data Hub can help you decide if an...

  25. Addgene's 20th Anniversary Party

    Type
    Blog Post
    Published
    June 11, 2024, 1:15 p.m.
    ...group of talented individuals that makes Addgene — well, Addgene. We're so grateful for each and every one...This year marks Addgene's 20th anniversary! We've been celebrating throughout the year, and on Sunday... to have many of our remote workers travel to Addgene's headquarters in Watertown, Massachusetts, for ...enjoying the party. There are so many teams that make Addgene functional, handling everything from facilities...  Throughout the party, we shared memories of Addgene's past. Framed photos and slideshows shared some...including an album of Addgenie pet photos. The famous Addgene jacket, home to a questionable collection of memorabilia...our co-ops to our directors, we can't celebrate Addgene without recognizing our incredible depositors and...

  26. Bringing Renewable Energy to Addgene

    Type
    Blog Post
    Published
    Aug. 27, 2019, 1:08 p.m.
    ....  Additional resources on the Addgene blog Subscribe to the Addgene blog Read our sustainability in...requires a significant energy expenditure here at Addgene. These are critical systems that cannot be removed...only sustainability effort we’ve implemented at Addgene. The kitchen and office areas provide multiple ... the lab as well. For a more in-depth look at Addgene’s current lab-oriented environmental efforts check... about conservation in the lab Resources on Addgene.org Sign up for our quarterly newsletter Help other...resources by sharing plasmids Find plasmids at Addgene ...

  27. It’s CRISPR Week at Addgene!

    Type
    Blog Post
    Published
    May 13, 2019, 12:01 p.m.
    ...social media. #AddgeneCRISPRweek on Twitter From May 13 (today!) to May 17, the Addgene Twitter feed will...CRISPR overview and history. Tuesday will focus on generating knockouts using CRISPR and Wednesday will highlight...CRISPR. Hope you will join us this week at #AddgeneCRISPRweek. We updated the CRISPR guide! If you’re not...

  28. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...HIV gag, pol, rev, and tet genes. It can be used with 2nd or 3rd generation lentiviral vectors and an ...3rd generation lentiviral packaging plasmid from the Trono lab that contains the gag and pol genes. It...packaging plasmid from the Trono lab contains the rev gene. It’s meant to be used with pMDLg/pRRE and the envelope...doxycycline to the grown media resulting in target gene knockdown. Note: this plasmid was called pLKO-Tet-On... 15 plasmids requested from Addgene psPAX2  - psPAX2 is a 2nd generation lentiviral packaging plasmid...their own or through Addgene!) pX330-U6-Chimeric_BB-CBh-hSpCas9 - The S. pyogenes Cas9 (SpCas9) has been...plasmid predates Addgene’s founding. Find pCMV-VSV-G. pRSV-Rev - This 3rd generation lentiviral packaging...

  29. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...reprogramming transgenes and the up-regulation of corresponding endogenous pluripotency genes. Retroviral...disrupt the expression of tumour suppressor genes and/or oncogenes, potentially leading to the presentation... Shao, L., & Wu, W. (2010). Gene-delivery systems for iPS cell generation. Expert opinion on biological...a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming factors... the transposase. The donor plasmid contains the gene(s) of interest flanked by terminal repeat sequences...demethylation and expression of endogenous pluripotency genes (i.e. Oct4) are other measures of reprogramming....pluripotency: history, mechanisms, and applications." Genes & Development 24, no. 20 (2010): 2239-263. PubMed...

  30. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    Published
    April 13, 2018, 12:51 p.m.
    ...Resources from the Addgene Blog Addgene's a nonprofit? Get an Inside Look at Addgene's NGS Quality Control... In this episode of the Addgene podcast, we sit down with Addgene’s Founders - Melina Fan, Ken Fan, and...interview with Addgene's founders Podcast breakdown Intro Chapter 1 - Who are Addgene's Founders? Chapter...Chapter 2 - The process of founding Addgene, where the idea for Addgene came from and how the idea got off...Chapter 6 - Developing Addgene’s Brand and culture Chapter 7 - Building Addgene as a global organization...current roles at Addgene and beyond - Ginkgo, Tufts Launchpad, and more Chapter 10 - Addgene's Future, the ...efforts at Addgene as the Chief Financial Officer and Chief Operating Officer. Prior to Addgene, Kenneth...

  31. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ... also modify other properties such as endogenous gene expression, which can lead to changes in normal ...neuronal firing. What is chemogenetics? Chemogenetics relies on genetically introducing engineered receptors... overview of the chemogenetics system. Why use chemogenetics Because chemogenetic receptors are activated...neuroscience Resources on addgene.org Learn more about chemogenetics in our chemogenetics guide Find viral preps...equipped using fast-acting optogenetic tools. The advantages provided by chemogenetics are instead that it is...longer-lasting effects. Building your chemogenetic system To set up a chemogenetic system, you’ll need to choose...most common target for chemogenetic manipulation are neurons where chemogenetic receptors can be used ...

  32. Addgene Launches Partnership with IPI

    Type
    Blog Post
    Published
    June 13, 2023, 1:15 p.m.
    ...Addgene is excited to announce the launch of our partnership with the Institute for Protein Innovation... of integrin antibodies, will be available on Addgene’s website this fall. IPI antibodies will come ... antibody sequences, in line with the rest of Addgene’s recombinant antibody collection, as well as access...developed to support use of their antibody tools. Addgene also offers a growing collection of educational...

  33. A Case Study of Addgene Bioinformatics

    Type
    Blog Post
    Published
    May 25, 2023, 1:15 p.m.
    ...collaboration.  In that post, we talked about Addgene’s data processing needs, our criteria for selecting...Recently, Seqera published a case study on how Addgene has used Nextflow to improve and redesign our pipeline...

  34. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...everything else we do! The goal of Addgene’s educational content Addgene truly believes that sharing speeds...you ever wondered how we choose topics for the Addgene blog? Or why we make certain videos? In this post...You probably won’t be surprised to learn that Addgene’s mission statement extends beyond plasmids, AAVs...topics. What better space to do that than in the Addgene blog?  Fig. 1: One of our favorite content...content collections is our Help Center, help.addgene.org, where you can easily search articles answering ...sure things are clear, accurate, and in line with Addgene style.        Fig. 2: Reference image (...already comes from you and the questions you ask Addgene and your research community! We work to provide...

  35. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ... the mutagenesis reaction. Alternative approaches to site directed mutagenesis Point-mutagenesis is fairly...relevant in this age of CRISPR; site-directed mutagenesis generally applies to plasmids, but may also facilitate...modifications.   Site-directed mutagenesis of Plasmids. A) Generation of mutagenized plasmids. PCR primers...also generates 4 bp overhangs. You can then use the Klenow fragment of DNA polymerase I to generate blunt...Series Use CRISPR to Generate Specific Genomic Mutations Resources on Addgene.org Browse Our Molecular...Laursen from Cornell University. Site directed mutagenesis is a highly versatile technique that can be used...analysis of proteins (to perform alanine scanning mutagenesis or targeted substitution of key residues), and...

  36. Introducing Addgene's Microbiology Pages

    Type
    Blog Post
    Published
    June 13, 2016, 7:08 p.m.
    ...work with fluorescent proteins, synthetic biology, gene cloning and expression, as well as guide pages for...plasmids made in your lab, see www.addgene.org/deposit or email deposit@addgene.org for more information or ...Addgene will be at the American Society for Microbiology Microbe meeting here in Boston next week - be... and say hello! Just in time for this meeting, Addgene is launching a new resource page for the Microbiology...organisms we should include by emailing us at help@addgene.org. If you are looking for plasmids used in E. .... coli and yeast, you’re in luck! Addgene has so many of these resources that we created filters just ...microbial technology, sign up to write about it on the Addgene blog. ...

  37. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ... caters to optogenetics users and adjusts the lasers appropriately (tuned for optogenetics). These lasers...answer a research question. The flip side of the optogenetics coin is materials science-based. Light is delivered...optics is probably the least difficult portion of optogenetics (though it does require a significant amount...become go-to providers of materials needed for optogenetics. There are several important considerations ...with fabrication of the fiber optics needed for optogenetics. First, the length of the fiber optic for the...laser output), 5ms “on”, 50ms “off”.  A pulse generator such as the A.M.P.I Master-9 Pulse Stimulator,...manipulation of the laser power source itself. Pulse generation (TTL pulse) is a standard electronics protocol...

  38. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...or negative selection). Next-generation sequencing (NGS) to identify genes of interest. Figure ...target genes for drug discovery or to understand the intricacies of molecular pathways. These screens have ...the targeting of hundreds to tens of thousands of genes with minimal effort. The versatility of Cas effectors.... (2022). AAV vectors: The Rubik’s cube of human gene therapy. Molecular Therapy, 30(12), 3515–3541. https... Adeno-associated virus as a delivery vector for gene therapy of human diseases. Signal Transduction and...Forward genetics screens are a valuable part of the molecular biology toolbox to identify new target ...lentiviral-based CRISPR libraries available at Addgene. The large packaging capacity and stable expression...

  39. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...provide an increased understanding of DNA binding and gene regulation. For additional information on CRISPR-mediated...technology using recombinant CRISPR ribonucleoproteins." Genes to Cells 21.4 (2016): 370-377. PubMed PMID: 26848818...interactions between genomic regions by enCh IP‐Seq." Genes to Cells22.6 (2017): 506-520. PubMed PMID: 28474362...the 3x FLAG‐dC as9 protein for enCh IP analysis." Genes to cells 23.4 (2018): 318-325. PubMed PMID: 29480524...RNA-guided platform for sequence-specific control of gene expression." Cell 152.5 (2013): 1173-1183. PubMed...Fujita et al., 2018). Furthermore, the Fujii Lab generated a transgenic mouse line and retroviral expression...specific gRNA-dCas9 complex. When combined with next-generation sequencing, this method allows for the recovery...

  40. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...-based genome engineering has revolutionized the gene editing field by making experimental workflows considerably... tools. As a consequence, a critical step in the gene editing workflow - analyzing the data - is often...likely to result in a functional KO of the targeted gene.   Analyze Complex Edits   Traditional Sanger sequencing-based...result in a functional Knockout (KO) of the targeted gene. The analysis can be sorted by any of the parameters...validation tool, and guaranteed editing with the CRISPR Gene Knockout Kit. Please contact us at [email protected]... more efficient than previous methods. Still, generating reliable results from CRISPR edit data requires...detect or analyze complex edits, such as those generated by multiplexed delivery of multiple sgRNAs. Since...
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