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We narrowed to 230 results for: cre encoding plasmid

Showing: 181 - 200 of 230 results

  1. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ... Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse Targeting...model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components are...genome is made up of 3 x 109 nucleotides (nt), and encodes 23,000 or so genes. It would be great if we could...target site, and a donor oligonucleotide or plasmid as the repair material if needed (for knock in models)...provided (in the form of an oligonucleotide or plasmid), precise changes can be made in the genome via...mice carrying the intended mutation. With a donor plasmid, the outcome is less predictable. In our best case...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...

  2. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...Non-coding RNAs (ncRNAs) are transcribed but do not encode proteins. They play important roles in many cellular...affect nearby genes (Figure 1A-C). Find CRISPR plasmids at Addgene! Knock-in: Introducing tags or stop...observe the effects. Luckily for us, CRISPR is an incredibly versatile system that can be tweaked to target...pairs long. Using a single guide RNA (sgRNA) to create a double-strand break works well because very small...ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for targeting thousands...092902 Liu Y, Cao Z, Wang Y, et al. Genome-wide screening for functional long noncoding RNAs in human cells...

  3. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

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    Blog Post
    ...sequence is altered but the protein sequence they encode is not. After each mutation, it will check the ...software Resources on Addgene.org Find CRISPR plasmids and resources Download the CRISPR eBook Read our...the sequence inserted by HDR may inadvertently re-create a match for the active guide. With these constraints.... To use this tool, simply follow these steps: Create a new experiment in the program Design gRNAs by...

  4. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

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    Blog Post
    ...Find recombinant antibodies at Addgene Find plasmids encoding recombinant antibodies at Addgene  ...epitopes previously hidden within the protein. Created with BioRender.   Permeabilization What is ...better at permeabilizing organellar membranes. Created with BioRender.   How do I choose?! Ok, so ...

  5. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...The most commonly used are GECIs - genetically encoded calcium indicators (e.g. GCaMP). Others include... comprehensive data sheets on many FP variants Plasmids 101: Green Fluorescent Protein (GFP) Three Review... the time it takes the FP to correctly fold and create the chromophore. This can be from a few minutes...

  6. CRISPR 101: Multiplex Expression of gRNAs

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    Blog Post
    ...assembled to produce a plasmid encoding the gRNAs and Cas9. As with the Gersbach lab plasmids, multiple Cas9 ...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...rectangles) are cloned into various plasmids using oligonucleotides. These plasmids contain Type IIS restriction...different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows you to express 2-4 gRNAs, with...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids can be used to express up to 8 gRNAs after...Yang Lab Single Transcript Multiplexing Plasmids: These plasmids are similar to the Csy4 polycistronic ...

  7. Antibodies 101: Introduction to Antibodies

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    Blog Post
    ...in cells that have been transfected with a plasmid encoding GFP. Actin, a common internal control for ... be used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies...immunoglobulin genes, recombinant antibodies are plasmid-based. Their sequences can be optimized for specificity...protein (Wang et al., 2013). These proteins are also plasmid-derived. However, a disadvantage to ScFv is their...nanobodies, or synbodies, can be produced from plasmids, saving time and money. Nanobodies and synbodies... as well as tools for developing and producing plasmid-based antibodies - and we’re always growing our...Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies for neuroscience research...

  8. Feng Zhang Multiplexed Overexpression of Regulatory Factors (MORF) Collection

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    Collection
    ... Zhang Lab Plasmids The Human TFome Library Stem Cells Protocols & References MORF Plasmids MORF Pooled...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...bulk screening (PDF, 85 KB) TF ORF barcode dial out PCR for 10X scRNA-seq (PDF, 59.2 KB) Plasmids ID Plasmid...transcription factor collection that contains 1,836 genes encoded by 3,548 isoforms and two control vectors. The ... Plasmid Gene/Insert Pooled Library The MORF pooled library is a comprehensive collection of human transcription...factor isoform in pooled screens. This collection and library can be used for screening transcription factors...available at Addgene. The collection can be used for screening transcription factors for cellular programming...

  9. TALEN Plasmids and Kits

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    Collection
    ...List of plasmid kits and accessory plasmids available from Addgene for generating your own specific TALE...Yamamoto Additional Plasmids for Use with Addgene TALEN Kits For Use With: ID Plasmid Depositing PI Description...in mammalian cells. These plasmids allow for cloning DNA fragments encoding TAL effector repeat arrays...TALE Toolbox 49622 - 49647 24 plasmids Thoru Pederson TALEColor plasmids: destination vectors for use ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...demethylase) made using these plasmids along with the FLASH method can decrease the amount of Histone H3 lysine... TALE plasmids. Genome...

  10. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ... the expression of a Cre recombinase contained within the knock-in. Including Cre-mediated marker excision...These modular toolkits build single plasmid targeting vectors that encode the gRNA transcript and the repair...kilobases of additional sequence being introduced. Cre-mediated excision of the selective cassette minimizes...experimental manipulation. In some cases, injection of Cre is required, however the development of the self-... C. elegans, visit Addgene’s C. elegans CRISPR plasmids page. Future perspectives It is interesting to...variable amounts of experimental manipulation and screening.  A co-selection strategy for CRISPR editing in... with inefficient sgRNAs in my initial direct screening efforts, which were similar to the approach detailed...

  11. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ... Blotting Resources on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...immunofluorescence and indirect immunofluorescence work. Image created with BioRender.com   Indirect IF is a much ...with key considerations about each step. Image created with BioRender.com   The first couple of steps...

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

    Type
    Blog Post
    ...BRDN0000561167) (Plasmid #80034) and BRAF gRNAs (BRDN0001145345 and BRDN0001146266) (Plasmid #76479 and #76480...was the most efficient for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702...viruses, but you can find a list of validated gRNA plasmids on our website. There are a few reasons to target...18007609. Addgene Materials Used 1. lentiCas9-Blast (Plasmid #52962) was a gift from Feng Zhang and is described... PubMed Central PMCID: PMC4486245.2. pXPR_011 (Plasmid #59702) was a gift from John Doench & David Root... MOIs. 2. Create a monoclonal Cas9-expression cell line. Especially if you’re doing a screen. I made a...who did not tell you that essential thing ask incredulously, “You seriously didn’t add 3 microliters of ...

  13. Special Delivery: Fluorophore Targeting for FRET Studies

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    Blog Post
    ...Resources on Addgene.org Find Plasmids for FRET Browse Empty Plasmids to Fluorescent Protein Fusions ...for genetically-encoded FRET donors (Fig. 1). Scores of FP variants have been created, thereby enabling...oocytes, mainly). Nevertheless, tRNA suppression plasmids are available from depositing labs that have mastered...the potential rewards could be substantial: Plasmid tRNA Synthetase Expression Host pMAH-POLY Polyspecific...Fig. 1) which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...are critical in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET...

  14. 27 Hot Plasmids from 2016

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    Blog Post
    ...Neurosci 2016. PubMed PMID: 26950006 Plasmid ID Plasmid Name Plasmid Type 74309 pcDNA3.0-TRPV4-p2A-ferritin-p2A-mCherry...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...features and applications of a partiular plasmid or set of plasmids. We hope that these articles make it ... you to find and use the plasmids you need. You can find all the hot plasmids from 2016 below. With over...over 45,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...stocks in a single plate with every plasmid or as individual plasmids shipped as agar stabs. Iverson, et...

  15. Chemogenetics Plasmids

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    Collection
    ... Plasmid Collections Chemogenetics Plasmids Chemogenetics Plasmid Collection You may...Chemogenetics vs. Optogenetics Chemogenetics plasmids encode genetically engineered receptors that are activated...Chemogenetics Plasmids Browse, sort, or search this table to find chemogenetics plasmids in our collection...ready-to-use AAV preparations of many chemogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...PSEMs). PSAMs are coupled to an ion pore domain to create a ligand-gated ion channel. Whether a PSAM is activating...

  16. Rinehart Lab Phosphoprotein Reagents

    Type
    Collection
    ...SepOTSλ Plasmid 68291 SepOTSκ Plasmid 68290 SepOTSι Plasmid 68289 SepOTSθ Plasmid 68288 SepOTSη Plasmid 68287...S218TAG Plasmid 68300 MBP-MEK1 Plasmid 68299 GFP E17TAG/Q157TAG Plasmid 68298 GFP S2TAG/E17TAG Plasmid 68297...GFP Q157TAG Plasmid 68296 GFP S2TAG Plasmid 68295 GFP E17TAG Plasmid 68294 SepOTSν Plasmid 68292 SepOTSλ...68287 SepOTSζ Plasmid 68286 SepOTSϵ Plasmid 68285 SepOTSδ Plasmid 68284 SepOTSγ Plasmid 68283 SepOTSβ Plasmid....ΔA.Δserb.Amp Plasmid 68305 Beta lactamase S68TAG Plasmid 68302 MBP-MEK1 S222TAG Plasmid 68301 MBP-MEK1...pSerOTS-C1* (V70) (Plasmid #188537) a better choice in place of the popular SepOTSλ (Plasmid #68292) . An important...-C1* (V70) is a ROP minus plasmid and is not compatible with other plasmids containing high copy ColE1...

  17. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...Blog Learn New Plasmid Cloning Techniques Pushing the Limits of DNA Assembly Plasmid Cloning by PCR ...including vector-insert joining for recombinant plasmid construction (restriction cloning), adaptor ligation...nucleotide sequence of a thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968...reliably detect single-base differences. The original creators of this technique visualized the ligation product...quencher pairs or qPCR-based methods to greatly increase the sensitivity of detection (14-20). LDR has ... 27). This methodology has been used to rapidly screen buffer conditions for Taq DNA Ligase and their ...electrophoresis (CE) (see below). This method allowed screening of all possible base combinations (Watson-Crick...

  18. Plan Your Experiment

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    Collection
    ...design. Addgene offers many plasmids containing validated gRNAs . These plasmids contain gRNAs that have ...cells is to directly transfect expression plasmids encoding your Cas protein and gRNA, which you generally...generate stable cell lines if your plasmid contains a selection marker. Plasmid delivery is versatile thanks...or mRNA encoding the Cas enzyme and gRNA, or as a functional ribonucleoprotein (RNP). Created with BioRender.com...Application Cas Enzyme Additional Considerations Browse Plasmids Knockout Permanently disrupt gene function; cells...different activator systems exist, including the multi-plasmid SAM system Activate Design and Clone Your gRNA ..., anneal, and insert the targeting oligos into plasmids containing the gRNA scaffold using standard restriction-ligation...

  19. A History of Genome Engineering in Popular Culture

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    Blog Post
    ... our CRISPR guide Find CRISPR plasmids for your research Browse plasmids for base editing References ...amino acids. Once scientists established that DNA encoded all the information necessary for regulating and...The premise of the novel involves Dr. Moreau who creates human-animal hybrids. This book is published 30...character revolts against class divide that is created by genetics.    Zinc finger nucleases Zinc finger... it can read many base pairs simultaneously to increase specificity. This modular approach made ZFNs highly...space travels. The movie sees the USS Enterprise crew fight against a genetically engineered superhuman...

  20. CRISPR Plasmids - Drosophila

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    Collection
    ...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...locus and encodes a primer binding site and the desired edits on an RT template. ID Plasmid Gene/Insert...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance... Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...CRISPR plasmids that have been designed for use in Drosophila and other insects. CRISPR...
Showing: 181 - 200 of 230 results