We narrowed to 1,147 results for: Gene

... we can make Addgene better and make scientists’ lives easier. Why transition to Addgene? So why did ...more satisfying, because Addgene is a non-profit, we can focus more on generating blog content that scientists... that PhD. Resources at Addgene Read Other Careers Posts Visit the Addgene Careers Page Learn about ...Harvard University and entered a new role here at Addgene as an Outreach Scientist. I used to spend my days... less time wrangling bacteria and instead help Addgene fulfill its mission of helping scientists share...did I join Addgene? Was I jaded by my time in academia? Did I run screaming from the lab like the cartoon...didn’t, it would have been unwise of me to join Addgene. This is particularly true in my position as an...

...domain due to a gene that resembles cas10. All type IV subtypes are missing adaptation genes, Cas1 and Cas2...sequence similarity, phylogenetic analysis, neighborhood analysis of surrounding genes, and experimental ...straightforward as it seems. There is not a single gene that is shared across all CRISPR types; the fast...together because they contain a distinct Cas7-type gene. The exact mechanism of type IV systems is not known... and applications with the type I CRISPR system. Gene and Genome Editing, 3–4, 100013. https://doi.org...the array of CRISPR technologies. Learning the general principles of this classification system will help...A–C, with the beloved SpCas9 (from Streptococcus pyogenes) belonging in II-A. Cas9 was the first to be adapted...

...’s first genetic engineering company, Genentech, was founded.  This commercialization of gene editing ...before the discovery of genes themselves.  How has scientific progress in genetic editing impacted the world... DNA sequences is not ideal. For the early generation gene editors, new amino acid recognition sites had...followed by non-homologous end-joining that results in gene knockouts.   Star Trek: Into Darkness (2013) which...The major drawback of all the endonuclease-based gene editors (meganucleases, ZFNs, TALENs, and CRISPR-Cas...Gregor Mendel on the genetic inheritance (1866) and just when the Mendelian genetics was being revived by...Zinc-Finger Nucleases. Genetics 188:773–782 . https://doi.org/10.1534/genetics.111.131433 Doudna JA, Charpentier...

...URL “http://n2t.net/addgene:87155” where the Addgene registered prefix and the Addgene ID # are directly...Daniel Bauer (Addgene plasmid # 87155 ; http://n2t.net/addgene:87155 ; RRID:Addgene_87155).” So when citing...Daniel Bauer (Addgene plasmid # 87155 ; http://n2t.net/addgene:87155 ; RRID:Addgene_87155)” Cite plasmid...through Addgene and find verified information about each plasmid item. Given a specific Addgene ID # listed...available through Addgene. Let’s take a look at lenti-Cas9-VQR-Blast, which has the Addgene ID #87155. The... as “RRID:Addgene_87155” and the following URL (https://scicrunch.org/resolver/RRID:Addgene_87155) will...contain the Addgene ID # and the information necessary to indicate that this ID is managed by Addgene. How to...

...when trying to edit a gene using homology directed repair, since HDR-mediated gene editing is most efficient...formerly Cpf1), a nuclease that generates double strand breaks in target genes resulting in the formation ...always positioned correctly to target a particular gene. Furthermore, a target sequence may have high homology...plasmids from this publication at Addgene. Additional Resources on the Addgene Blog  Read more about Cpf1...sequences While PAM sequences for the commonly used S. pyogenes Cas9 (3'-NGG) are abundant throughout the human...circumvent this limitation: 1) the use of novel S. pyogenes Cas9 variants with varying PAM sequences, 2) the...Cas9 homologs derived from species other than S. pyogenes, and 3) the use of non-Cas9 enzymes. (For more...

...edits of interest than solely transfected cells. “Gene On” (GO)- a functional reporter system to identify... (Katti et al., 2020). They named this reporter “Gene On” or GO. GO works by affecting protein translation...fluorescent proteins. Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter...and enrich for base-editing events in vivo. TLDR: Gene On (GO) from the Dow lab is a flexible reporter ...base-pair mCherry insertion. Editing of these motifs generates lesions that are subject to excision and ssDNA...This edit changes the histidine to a tyrosine generating a GFP variant (GFPY66) that has a shifted emission...hypothesis with GFP fluorescence. To do this they generated a silent GFP construct, that contains an ACG start...

... maintain the genes they carry among organisms belonging to many different phylogenetic groups (Jain and...that the plasmid is conjugative and carries the genes necessary for transfer initiation between bacterial...plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197. https://doi.org/10.1016/0378-1119... S. (2011). Broad-host-range plasmid vectors for gene expression in bacteria. Methods in Molecular Biology...viscosus, Aerobacter aerogenes, Aeromonas hydrophila, Agrobacterium tumefaciens, Alcaligenes eutrophus, Azotobacter...salmonicida, Agrobacterium tumefaciens, A. rhizogenes, Alcaligenes eutrophus, Enterobacter sp., Erwinia amylovora... broad host-range plasmid RK2. Molecular & General Genetics: MGG, 152(3), 129–135. https://doi.org/10.1007...

...homology arm as a gBlocks® Gene Fragment from Integrated DNA technologies. Generally speaking, anyone using... an effort to track down every last one of those gene regulatory proteins and the places in the genome...together on several projects and this is one related to gene expression. One goal of the ENCODE project is to...with detailed protocols, go to www.addgene.org/crispr/tagging. Addgene: What motivated you to adapt CRISPR...tagging system, which they’ve now made available at Addgene. For more details on the plasmids that are currently...can now look at many more transcription factors. Addgene: How does the system work? Mendenhall: The system...antibodies. The results look almost identical. Addgene: What advantages does your system have in this ...

... to turn on genes at different times. It requires more complex synthetic networks. Addgene: What hurdles...on an engineering project including hundreds of genes and regulation and resource control - everything...gives us enough hardware to control hundreds of genes. We’ve created enough regulatory elements, CRISPRs...the scale of genetic engineering, with the ultimate objective of genome design.” It’s genetic engineering...larger genetic circuits that could conceivably be as large as natural regulatory networks. Addgene: What...things are easy with CRISPR. Addgene: You’ve made many deposits to Addgene over the years. Can you describe...toolbox for taking on very large genetic engineering projects. Addgene: How easy is it to do this now? ...

...the complexity of genetic circuits, allowing for tighter spatio-temporal control of gene expression within...world of possibilities in the engineering of complex gene networks. A comparable system for targeting specific...will make those new plasmids available through Addgene, and they hope others will do the same. “This isn...10.1186/1754-1611-8-7. PubMed.  Related Resources Addgene's Synthetic Biology Overview More Synbio Blog Posts...

...in neurons. (b) Genetic cargo can be customized to control transgene expression. Gene regulatory elements....     Figure 1: AAV toolkit for targeted gene delivery to the brain. The AAV (a) capsid, (b) cargo...Abbreviations: AAV, adeno-associated virus; GRE, gene regulatory element; NHP, non-human primate. ...LY6C1, with relatively even distribution of target gene expression (EGFP, in the depositing manuscript) ...Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature Protocols;14, 379–414...2017). Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems...Gradinaru, V. (2022). Engineered AAVs for non-invasive gene delivery to rodent and non-human primate nervous...

...looks like the genetic cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate...with those particles. Since the Env, Gag, and Pol genes are not encoded in the transfer plasmid, the component...Similarly, if the viral vector carries an shRNA to a gene essential for cellular function, this would also...Resources Resources on Addgene.org Addgene viral services Addgene Biosafety Guide Addgene Viral Plasmids ...? That’s okay! In this article we will discuss general viral safety considerations and best practices ...of what is being integrated by the virus. If an oncogene overexpression construct is being delivered, that... and the same holds true for types of viruses generated in the lab. The four most common types of viruses...

...existing genes, while being accessible simply by dialing up the right nucleic acid sequence (Addgene can help...DogCatcher); ideal for protein loop regions CRISPaint Gene Tagging Kit SpyTag/SpyCatcher CRISPR-assisted insertion...these systems can also assemble vaccines, target gene therapy vectors, or lock together smart biomaterials...handful of Click Biology tools available at Addgene. Addgene Link Click Biology Tag Description Humberg...of us still do), so we are used to the idea of general interfaces and the freedom to test out lots of ...one finds in cells. High yield of reaction — generating product without a lot of left-over starting components...possible through the utilization and development of genetically engineered components, like protein tags (Figure...

...Additional Resources on the Addgene Blog Treating Muscular Dystrophy with CRISPR Gene Editing Learn to Use ...Cas9 have also been used to create mutations in genes essential to HIV-1 replication. However, this approach...modification targeting either the LTRs or essential genes. Many of these escape mutations were located near...Genomes from Human T-lymphoid Cells by CRISPR/Cas9 Gene Editing. Sci Rep. 2016 Mar 4;6:22555. PubMed PMID...Screens Additional Resources onf Addgene.org Check out Addgene’s newly updated CRISPR Guide! Find CRISPR...Activation Mediators (SAM) system available from Addgene. This system uses the dCas9-VP64 fusion commonly...others and accelerate the pace of research. We at Addgene hope to see the problem of in vivo CRISPR delivery...

...ultrasound to image bacteria in vivo: acoustic reporter genes’ on our blog. Creative approaches to presenting...Bourdeau, Raymond W., et al. "Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian...way to visualize them. You can also “hear” them. Addgene depositor Dr. Mikhail Shapiro from Caltech presented...loads of inspiration for the current and the next generation of microbiologists. If you got inspired to produce...PubMed Central PMCID: PMC5920530.  Resources on the Addgene Blog Read Our Post on CRISPR Methods for Bacterial...Science Communication using Twitter  Resources on Addgene.org Check out Our Microbiology Resource Page CRISPR...bacteria Deposit Your Microbiology Plasmids with Addgene ...

...new green glowing protein could be used to measure gene transcription, as well as to track protein localization... he was right! He began to study the A. victoria gene responsible for encoding GFP, and, in 1992, he reported...

...Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids from the...treat disease through tissue repair, stem cells, gene therapy, and tissue engineering. How the multidisciplinary...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School...source biology in his own lab, at the Center for Regenerative Medicine, and within the biomedical research...Along these lines, Darrell recently joined the Addgene Board of Directors where he hopes to help keep ... Kotton and the Boston University Center for Regenerative Medicine. Darrell describes his career goals...

...disease-causing alleles, and the growing need for gene editing in research, the old PAM constraints just...RNA will do. Similarly, if you want to knock out a gene, a frameshift mutation introduced anywhere within...take a bp or two upstream or downstream) for most gene targeting experiments. Between SpRY and several ...Resources on Addgene.org Addgene’s CRISPR Guide CRISPR Plasmids and Resources Resources on the Addgene blog ...biology, the answer is always it depends, but generally the efficiency of the desired edit is reduced ... Cas9s that recognize NG PAMs. The Nureki lab generated SpCas9-NG, a rationally engineered Cas9 with NG...name implies (Nishimasu et. al, 2018). xCas9 was generated by the Liu lab through phage-assisted evolution...

...Plasmids for endogenous gene tagging in human cells Resources on Addgene.org Browse all CRISPR plasmids...endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and dTAG-13 ...endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, dTAG-13, and...Figure 1). Plasmids: We generated a series of plasmids that are available from Addgene. These plasmids enable...47 Additional Resources on the Addgene Blog Easi-CRISPR: Generating Knock-In and Conditional Mouse Models.... To overcome this limitation, we created a generalizable tag-based strategy, which we refer to as the... scales that are not possible with traditional genetic approaches. In addition, the dTAG system is versatile...

...overexpressing a gene or protein after transfection). Future of FRET The first genetically encoded FRET biosensor...Resource page and can be used to create fusions with a gene of interest. These plasmids will be most useful ...designed to measure specific small biomolecules or gene activity are often intramolecular probes, as the...name a few.   Find Many FRET-Based Biosensors on Addgene's Biosensor Pages   Recent innovations have demonstrated...PubMed PMID: 26210398. Additional Resources on the Addgene Blog Tips for using FRET in Your Experiments Learn...FRET Based Biosensors Additional Resources on Addgene.org Find FRET Pairs Check Out Fluorescent Protein...