...several candidate inhibitors of gene X in a screen. You generate a gene X knockout in a cancerous background...inhibit, activate, or knockout many genes at a time to identify ‘hits’ - genes that affect disease progression...some of the candidate genes! Disease model generation CRISPR has made the generation of genome edits accessible.... These hits can reveal individual genes or sometimes even genetic pathways that are worth investigating...known – repeat expansion, gene knock out, etc. – CRISPR can be used to generate cell line or animal models...resources on the Addgene blog CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease...approaches. The first route involves identifying a gene/pathway to develop a drug for (aka a druggable target...

...the Grousbeck Family Chair in Gene Therapy, Director of the Grousbeck Gene Therapy Center, Associate Director...at KSQ Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test...nonprofit companies. His lab focuses on gene therapy work for genetic diseases and over the past few months...deposited AAV constructs for gene delivery. In 2018, he interviewed Addgene co-founder Melina Fan and former...PhD Khandan Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development,...Dimension Therapeutics, she helped build and grow the gene therapy division of the company.  Ultragenyx’s goal...test the function of all 20,000 genes in disease models and then focus drug discovery efforts on the targets...

...repression of your target gene, activation and repression of different genes in a single cell, and localization...replication and transcription as well as reporter genes. To learn how you can use minigenomes in your research... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (... (AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “...localization of a fluorescent signal to your target gene. Read this post to find out more about how the Casilio...are widely used by the optogenetics and chemogenetics communities to activate neurons and neuronal pathways...

...in applying gene technology to bacteria and plants. Her research involves cloning cry genes from the soil...plant, Dr. Robertson aims to make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy... shared mission of both Seeding Labs and Addgene. The Addgene-Seeding Labs plasmid grant is a collaborative...of the Addgene-Seeding Labs Plasmid Grant awardees' projects progress. Subscribe to the Addgene blog for.... Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant Check...introduce the 5 scientists who were awarded the Addgene-Seeding Labs Plasmid grant this year. These scientists...plant biology, she coordinates the Network for the Genetic Improvement of Cowpea, a knowledge database for...

...lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1...specifically investigated protein binding at the TERT gene, but this method enables the unbiased discovery ...cDNA sequencing and RNA sequencing of complex heterogeneous RNA samples. Furthermore, such high processivity...genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase, (APEX)...are enriched in the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology... technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements by restricted spatial...

...Using CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org Find Plant ...experiments outlined in ABRC’s kits vary by module. As a general guideline, experiments that include growing plants...-12 education.     Additional Resources on the Addgene Blog Learn how BioBuilder can bring Synthetic ...Plant Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants Parts Kit ...

... delete/overexpress endogenous genes or to introduce heterologous genes, but the trend of research has...) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...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...

...systems allow researchers to understand bacterial gene clusters that produce useful natural materials, ...capable of robust expression of high G+C content genes, as validated through monitoring the expression ...end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter for editing...human mRNA binding proteins with 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode...pooled library from Junjie Chen's lab targets human genes known or suspected to be involved in DNA damage ...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons in ...library uses CRISPR cytosine base editors to disrupt gene function by targeting start codons, splice acceptor...

...journal club discussion on Kevin Esvelt’s CRISPR/Cas9 gene drive paper was very popular. People kept asking... at Addgene.org Order Plasmids from Addgene! Find Free and Open Educational Resources at Addgene! ...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...microbes. We’ve ordered a few yeast plasmids from Addgene and are eager to do our first yeast transformations... @ZebraElement     Additional Resources on the Addgene Blog Experimenting in an Open Source Lab: from...

... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...Fatima's work and keep checking back in with the Addgene blog for future award opportunities. Xin Zhou:...when exposed to ultraviolet light. To do so, she generated fusion proteins in which Dronpa domains flanked...Proteins, or FLiPs.  Xin later optimized these first generation FLiPs by mutating Dronpa to lower its aggregation...Chemical Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent...Fluorescent Protein for Your Experiments Resources on Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids...

... transcription of a target gene, it therefore determines the timing of gene expression and largely defines...activating transcription. Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering...synthesis and allowing reporter gene transcription to take place. Addgene also has yellow flame 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...

...all known genes. This is a perfect example of an unbiased, reverse screen – all known genes are assayed...of knocking out the genes of interest (or the whole genome), they knock down gene expression temporarily...Sabatini, D.M., Lander, E.S. Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166... a biological process. Players can be proteins, genes, enzymes, drugs, or anything else that plays a role...where there is removal of known factors (such as a genes) and the loss of those factors is assessed. The ...especially useful if you are interested in studying genes whose knockout would be lethal to cells and thus...The two most common methods of manipulation are genetic knockout mutations and treatment with small molecules...

...used with many types of proteins, including Cas9 and dCas9-transcription factors, for which low expression...plasmids from this publication at Addgene. Multichromatic control of gene expression in Escherichia coli...optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...used to engineer new pathways with sophisticated gene expression controls. Most cell types do not respond... can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive two-component... was only about 10-fold, precluding the study of gene expression at very high or very low levels. Schmidl...researchers to better fine-tune the level of output gene expression based on the input light intensity. In...

...from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation and activation... popular tools for genetic screens, enabling genome-wide interrogation of the genes required for cellular...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 ...

...ampicillin resistance gene, you’ll find the full name of the gene, the name of its gene product, and a description...features, such as Cas9, weren’t detected by our in-house software. In fact, SnapGene’s feature library ... 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...

...currently available to industry scientists. ID Plasmid Gene/Insert PI Kits Kit name Type PI Description High...Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing of human TP53...Packaging from Addgene Parkinson's Research from The Michael J Fox Foundation Search Use Addgene's Search to...protection for material providers. Addgene’s deposit agreements allow Addgene to distribute plasmids on behalf...can be submitted to Addgene through your user account or by email to mta@addgene.org . After we review ...A portion of Addgene’s collection is available to qualified for-profit entities. Addgene reserves the ...Addgene has a growing collection of plasmids that are available to industry. Learn more about the available...

...changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the...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 (...

... delivery of CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids...Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential strategy is to...applications in diseases which may be linked to epigenetic changes, such as multiple sclerosis?  A: Basic...an exciting area for future AAV application to epigenetic editing. This area is discussed in our recent...excellent questions.    Additional Resources on the Addgene Blog Using virus in your research - a primer for...

...eNLs as good reporter genes for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Learn about Photoactivatable...proteins. To make luminescent imaging a reality, Addgene depositor Takeharu Nagai and colleagues at Osaka...nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion proteins. Solving the...yield, and when transiently expressed in cells, generates much less signal than comparable fluorescent proteins... approach is especially suited to complement optogenetic tools, where light used for excitation can cause...cause unintended activation of the optogenetic system. Biosensors based on Nano-lantern technology are...

...situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even here...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...PMCID: PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which... tagging strategies Additional Resources on Addgene.org Fluorescent protein plasmids Use FRET to study...protein-protein interactions Deposit plasmids with Addgene! ...