We narrowed to 1,141 results for: Gene
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TypeBlog Post...detect gene copy number based on the number of fluorescent puncta observed. By labeling two genes of a ...vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing, but it retains its gRNA-directed targeting...Chen et al. successfully labeled protein-coding genes with both intronic and exonic gRNAs. In fact, the...also monitor the spatial relationship of the two genes over time, at a range of distances from 2-75 Mbp... envision using CRISPRainbow in combination with gene editing. CRISPRainbow requires very short, 11-mer...plasmids from this publication at Addgene. Additional Resources on the Addgene Blog Read More about Fluorescent...cells! This technique was developed in the lab of Addgene depositor Stanley Qi and used in Chen et al. to...
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Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening
TypeBlog Post...functionally characterize combinatorial gene knockouts generated via multiplexed gRNA expression (Wong ...et al., 2014; Konermann et al., 2014) individual genes for studies interrogating their functions. The continual...list of effective gRNA sequences targeting your genes of interest. Thanks to the tremendous efforts made... et al. "Genome-scale CRISPR-mediated control of gene repression and activation." Cell 159.3 (2014): 647... "High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities." Cell ...Hongming, et al. "A CRISPR-based screen identifies genes essential for West-Nile-virus-induced cell death...incredibly useful for the efficient generation of individual (and multiple) genetic perturbations in single cells... -
Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP
TypeBlog Post...label cells. If GFP could be co-opted to control gene expression, he could then selectively manipulate...used LexA and rTetR systems such that they activate gene expression from different promoters. Moving to an.... GFP expression robustly activated the reporter gene TdTomato, whose expression was absent without electroporated...sites. Placing a lox-STOP-lox cassette upstream of a gene of interest blocks transcription when Cre is absent...present, it removes the STOP cassette, activating gene expression. Although Cre isn’t a modular protein...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed...commonly used in optogenetics, opening up the possibility of adapting GFP lines for optogenetics experiments... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog Post...deletion of one or several of these genes permits the expression of genes farther downstream in the construct... two tandem sets of genes aligned head-to-head and the deletion of one of the gene pairs. Brainbow 2.0...Brainbow uses a Cre/lox system to label neurons. Genes encoding the fluorescent proteins are organized ...organized in tandem with a single promoter, but only the gene directly following the promoter is expressed. Cre-mediated... which uses Cre to trigger the inversion of two genes aligned head-to-head. Another variation, Brainbow...located directly upstream of the fluorescent protein genes, Cre recombination is required before fluorescence...fluorescent proteins topic page Resources on Addgene.org Browse Addgene’s fluorescent protein collection Learn... -
Five Popular Model Organisms, Part 2
TypeBlog Post...human genes have counterparts in the rat (Gibbs, et al., 2004). Additionally, breakthroughs in gene modification...abundance of genes related to limb regeneration (Vieira et al., 2019). To study regeneration scientists ...during and after embryogenesis. Addgene has over 600 plasmids that contain Xenopus genes and a tool-kit ...models. Check out Addgene’s collection of plasmids containing Rattus norvegicus genes! Mexican axolotl ...array of molecular tools, such as gene manipulation and mutagenesis techniques, are available to the scientific...world worked to characterize each gene’s function. Some key genetics research with Arabidopsis include... for scientists to study human disease-relevant genes in such a controlled environment displays how essential... -
Plasmids 101: Restriction Cloning
TypeBlog Post...you were cloning a gene into an expression vector, you would want the start of the gene to end up just downstream... 5’ of the start of the gene and a HindIII site just 3’ of the end of the gene. This way you can then ...leaving the 5’ end of your gene just downstream of the promoter and placing the gene in the proper orientation... multiple sites allows you to easily orient your gene insert with respect to the promoter. For example...restriction sites EcoRI, XhoI, and HindIII. To place your gene in the proper orientation downstream of the promoter...binds the plasmid to the new fragment thereby generating a complete, circular plasmid that can be easily...green, insert) are cut with restriction enzymes to generate compatible overhangs that allow them to bind. ... -
Your Top Requested Plasmid in 2016!
TypeBlog Post...the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables selection of cells...to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone...had a point mutation in the puromycin resistance gene that made it less effective in certain cell lines...Resources on the Addgene Blog Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read...Purpose SpCas9 This plasmid expresses Streptococcus pyogenes (Sp) Cas9 from a Cbh promoter. Cbh is a modified... the key component of most CRISPR systems and generates double strange breaks at target sites 5' of the...your 20 nucleotide guide sequence of choice (for generation of a gRNA that will target SpCas9 to the approrpriate... -
Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative
TypeBlog Post...the sgRNAiBAR library covers every annotated human gene, it enables genome-wide screening with a high MOI...a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed, the...fish, and mice, by replacing the effector with a gene of interest in the cells you are studying. Read ...calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin is required for ...antifungals that target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay,...Competition (SPARC) is a fully genetic tool for labeling just a fraction of a genetically-defined subset of cells...lentiviral vectors the iBARs can be detected by next generation sequencing. In order to avoid a high false-positive... -
CRISPR References and Information
TypeCollection...identify positively or negatively selected sgRNAs, genes, or pathways. For use with genome-scale CRISPR-Cas9...CRISPR screens targeting a long or short list of genes provided by the user. Developed by the Fredrik Wermeling...sgRNA design tool for genome editing, as well as gene regulation (repression and activation). Genome support...target sites by off-target quality. Validated for gene inactivation, NHEJ, and HDR. Includes a wide range... to identify novel gRNA target sites in a single gene, as well as a target site common to a set of similar... Information Addgene Resources CRISPR Software General Resources CRISPR Protocols Addgene CRISPR Resources...Resources CRISPR Plasmids Addgene's collection of CRISPR plasmids. CRISPR Guide Addgene's guide to using CRISPR... -
Feng Zhang Multiplexed Overexpression of Regulatory Factors (MORF) Collection
TypeCollection...transcription factor collection that contains 1,836 genes encoded by 3,548 isoforms and two control vectors...10X scRNA-seq (PDF, 59.2 KB) Plasmids ID Plasmid Gene/Insert Pooled Library The MORF pooled library is... transcription factor collection, available at Addgene. The collection can be used for screening transcription... entire pooled collection, can be purchased at Addgene. Each transcription factor isoform has a unique...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog Post...sensitizing gene cassettes. Thse vectors are engineered to replace the drug resistantance genes in the bacteria...were identified for 61 such diseases. Now single gene alleles have been identified for 4,163, over 50%...Lowe talked about his work on the 500+ human tRNA genes and, until now, we've overlooked regulatory fragments...may be involved in preventing silencing of self-genes. Dr. Mello presented a hypothesis that these pathways...involved in RNA based surveillance of repetitive gene families and provide a way to guide inheritance ... field. Resources at Addgene Browse Addgene's RNAi resources Browse Addgene's CRISPR resources Read ...countries in number of plasmid requests from Addgene. Addgene - Canton Nucleic Acids Forum 2015 from Joanne... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog Post...Ehrhardt, A., et al., Episomal vectors for gene therapy. Curr Gene Ther, 2008. 8(3): p. 147-61. PubMed PMID... functional genes at the site of insertion and which can potentially promote tumorigenesis in vivo. The...fellow at Massachusetts General Hospital with research interests in the fields of gene editing and stem cells...non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res, 2011. 21...The phenomenon has been recorded in past gene therapy trials and remains a safety concern in the clinical...clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science, 2003. 302(5644): p...the chances of insertional mutagenesis, which is the integration of transgenes into the genome that can ... -
Plasmids for Stem Cell Research
TypeCollection...table based on plasmid name, species, gene name, and more. Plasmid Gene/Insert Vector Type PI Publication ...cell lines with endogenously-tagged gene variants or browse Addgene’s entire collection of Stem Cell Research...against p53 in different gene/insert combinations A more efficient method to generate integration-free human...non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res. 2011 Mar...human cells to pluripotency by activating enogenous genes Human pluripotent reprogramming with CRISPR activators...S1934-5909(19)30423-0. Schöler You can also find gene-specific plasmids at the following links: NANOG ...Browse Addgene's collection of plasmids for the generation and study of stem cells, from iPSC generation... -
A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog Post...editing single genes is very useful, sometimes, you need to modify multiple genes to generate a cellular ...Nicole Gaudelli, the Associate Director and Head of Gene Editing Technology at Beam Therapeutics, used a ...In addition to stem cells the authors targeted 6 genes in primary human T cells using 8 selected ABE8s ...the reduction in protein expression of these six genes. All ABE8s edited more efficiently than ABE7.10 ... was the winner. It could simultaneously edit 3 genes (98.1%, 98.3%, or 98.6% efficiency, respectively... eighth generation of adenine base editors, called ABE8 (Gaudelli et al., 2020). The generated ABE8s needed...replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog Post...used for genome editing, activating gene expression, repressing gene expression, and much more. But there...engineering viruses. For example, an Acr gene was used to replace genes in the difficult-to-engineer Sulfolobus... Selection marker while engineering viruses Acr genes can be used as positive selectable markers in engineering...et al., 2018). Only viral particles that have the gene deletion could replicate when challenged with the...Experiment Read more about Optogenetic Control of CRISPR Resources on Addgene.org Find CRISPR Plasmids ...Nme) (Pawluk et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and well studied...10.1126/sciadv.1701620 Additional Resources on the Addgene Blog First time CRISPR user? Check out this post... -
Anatomy of a Plasmid Page at Addgene
TypeBlog Post...within the gene/insert, this information can be found here. There may be more than one gene inserted into...can easily clone my gene into it? Your best bet is the Plasmid Map: We auto-generate a plasmid map based... deposited with Addgene, you can list the original vector’s plasmid number here. Gene/insert: This describes...the backbone, and provides information about this gene or DNA element of interest. If the depositor has... into a backbone, and each is given its own gene/insert section on the plasmid page. Tags are listed according...We Do Quality Control at Addgene Resources on Addgene.org Join the Addgene Community Learn the Benefits...plasmid. Use the Addgene catalog number to easily find template citations for Addgene plasmids you have... -
Allen Institute for Cell Science Plasmid Collection
TypeCollection... Endogenous Gene Tagging in Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human...Link opens in a new window) RNP transfection for gene editing human iPS cells (Link opens in a new window... The plasmids used to generate these lines have been deposited here with Addgene for distribution to the...Institute for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular ...With these plasmids researchers will be able to generate fluorescently tagged proteins of key cellular ... with endogenously-tagged cellular structures. Addgene ID Plasmid Allen Institute ID Tag Protein Structure... -
Optogenetics Plasmid Collection
TypeCollection... AAV preparations of many optogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter..., "Hahn", or other terms. ID Plasmid Description Gene/Insert Vector Type Promoter PI Publication Return...Collections Optogenetics Plasmids Optogenetics Plasmid Collection You may also like... Optogenetics Guide AAV...tables, refer to Addgene's Optogenetics Guide . Interested in more news about optogenetics research? Find... AAV Viral Service Biosensors Chemogenetics Plasmids Chemogenetics Guide Fluorescent Protein Guide Microbial...Find optogenetics plasmids such as microbial opsin and optical switch tools to measure and manipulate... Plasmids Optical Switch Plasmids Background Optogenetics plasmids encode proteins that are controlled... -
CRISPR Plasmids - gRNAs
TypeCollection... lists gRNA plasmids designed to target various genes or genomic regions. It is highly recommended that...the experimental use of these plasmids. ID Plasmid Gene/Insert Vector Type PI Publication Do you have suggestions...gRNA expression vectors available from Addgene. CRISPR...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other... effect. Which species or variant of Cas9 ( S. pyogenes , S. aureus etc.) was this gRNA sequence designed...promoters. Do you have gRNAs you'd like to add to the Addgene collection? Click here to start the deposit process...plasmids added to the list. You can also email help@addgene.org for more information and assistance with the ... -
CRISPR Plasmids - CRISPR Transposases (CAST)
TypeCollection... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Selectable...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...out our Suggest a Plasmid form or e-mail help@addgene.org to help us improve this resource!...