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We narrowed to 956 results for: REP

Showing: 461 - 480 of 956 results

  1. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...ADAR constructs from the REPAIR (RNA Editing for Programmable A to I Replacement) system. Like RNA targeting...Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed Repair and 3 Tips to Improve HDR Efficiency...Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair mechanisms: CRISPR 101: Non-homologous End Joining...

  2. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ... Specifically, viruses are able to infect and replicate in host cells, but they are susceptible to cells...transposon consists of a mirrored set of inverted repeats (green) flanking the gene of interest (blue) in...is expressed (red stars) and binds the inverted repeats (green); and an endonuclease reaction occurs which...puromycin, hygromycin) and fluorescent protein reporters (GFP, BFP, RFP, no FP). The vectors each contain...region of the RPL41 gene, to drive the selection/reporter parts of each construct. This new tool kit allows...

  3. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...plasmid. Inverted terminal repeat (ITR) sequence Inverted terminal repeats (ITRs) are what makes an AAV...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...’s ~4.4 kb or smaller. Some examples include a reporter like GFP, some smaller CRISPR-Cas proteins such...(2019) A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated Virus. Human Gene Therapy Methods...

  4. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    .... In doing so, the dimer shifts from one that represses transcription to one that activates transcription.However...BLADE The team modified AraC to create BLADE by replacing its dimerization domain with VVD, a light-responsive...the DNA-binding domain of AraC using a mCherry reporter downstream of a PBAD promoter. While none of the...contains BLADE, the PBAD promoter, and a mCherry reporter downstream of the PBAD promoter. To use this plasmid... plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY:...

  5. What the HEK?

    Type
    Blog Post
    ... This allows them to replicate viral plasmids with the SV40 origin of replication. To generate a virus...considered hypotriploid: most chromosomes are represented by two to three copies, but several have four...your research question, HEK-293 is known for reproducible results. It’s also widely available, which can...biology, ion channels, or involve human protein preparations with PTMs. Why the HEK not? References and ...

  6. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    ... posts of 2015 1. CRISPR 101: Homology Directed Repair Addgenie Chari Cortez explains some of the basic...basic science behind homology directed repair (HDR) and how it’s used in CRISPR genome editing.   2. Plasmids...While the vast majority of CRISPR plasmids in our repository use Cas9 to cut DNA, Addgene Science Communications...explain some of the molecular gymnastics behind DNA repair by non-homologous end joining (NHEJ). 7. Career...Resources Learn More about Viral Vectors Browse the Repository  ...

  7. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ...You’ve prepped your DNA and you’re ready to get started on the next step of your experiment. But in many...give you an idea of the concentration of your DNA prep and whether there are any other contaminants. Nucleic...your sample against this curve to quantify your DNA prep. Though this method takes more time to set up in...and is not used as much for the standard plasmid prep. Diphenylamine method Another absorbance based method...Learn about the different grades of plasmid DNA preps Resources on Addgene.org Visit the Molecular Biology...

  8. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    ... about sexual and reproductive health, as the stigma around sexual and reproductive health issues creates...to all the optional sexual health sessions and reported the intel back to my shyer classmates. For my ...evaluate the quality of the study those abstracts represented. The point of this study was to see if their ...research focuses on misinformation about sexual and reproductive health.    References and Resources References...

  9. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...Gerald Schwank, first author of the cystic fibrosis report. “This is a big advantage over induced pluripotent...also need to be perfected until the efficiency of repair approaches 100 percent. “We need to be sure it'...it's accurate enough and efficient enough for repair,” Doudna said. “And it can't be too hard to apply....need to get a handle on how cleavage sites are repaired.” Those insights will be critical for scientists...December 2013). 8. Gerald Schwank et al. "Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell ...

  10. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...enough to get it into cells and let them do the repair work. The key is T4 DNA polymerase, which has 3...forming a circular product with four nicks easily repairable by the bacteria after transformation. LIC is ...transformed into bacteria, which will efficiently repair the plasmid. Created with BioRender.com.   ...intermediate is transformed into bacteria, which repair the plasmid. Created with BioRender.com. How...PCR-generated inserts. By harnessing the power of DNA repair, you can assemble multiple fragments without the...

  11. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    ...convinced that most microscopy techniques can be replicated with commercially available consumer-grade components... tools has allowed the community to regularly replicate designs worldwide. Distributing hardware via globally...Detailed documentation, which facilitates the reproduction of the structure in a similar way to the biological...assemblies open-source. The sharing of digital representations of the 3D models, which in many cases can be...with the goal of creating more transparent and reproducible science in the long run (Hohlbein, et. al., ...

  12. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...plays an integral role in regulating genome replication, repairing mismatched basepairs or small indels that...occur during DNA synthesis, and protomoting or repressing protein expression. Methylases involved with ... (as these would also be deficient in mis-match repair functions of Dam), so these strains should not ...Learn to Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources ...

  13. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...knockout screening). As the Zhang lab's Nature paper reports, “Here we have shown that the SAM system is robust...investigation of the cellular consequences of repressing or inducing individual transcripts. The activator...site in 15,977 human genes. The transcriptional repressor (CRISPRi) library contains 10 sgRNAs for each ...be turned on and off, to reversibly activate or repress genes of interest. They also “showed that different... Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson...

  14. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...proteins known as ankyrin repeat proteins. DARPins typically consist of 3-4 repeats, have high specificity...instead of generated in vivo, they have higher reproducibility and consistency in the final product. Because...refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically engineered...combinatorial libraries, and high specificity has been reported for a number of targets. Due to their size and...

  15. Antibodies 101: Secondary Antibodies

    Type
    Blog Post
    ...Primary antibodies are represented in blue; secondary antibodies are represented in purple; signaling molecules... molecules are represented in light green; and proteins are represented in orange. Created with BioRender.com...architecture of the indirect approach is as follows: Prepare the sample. Incubate the sample with the primary...

  16. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...catalytically dead Cas9 fused to a transcriptional repressor to your gene of interest. It will inhibit transcription...more on this below) will both achieve temporary repression of protein expression. Addgene has a dCas9 KRAB...CRISPRa blog! Fig. 1 CRISPR activator and repressor mechanisms Want to transiently induce expression...suited for the job. Whether you want to express or repress, good luck on your protein expression journey! ...Plasmid 101: Inducible Promoters Plasmid 101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose...

  17. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...the crRNA also contains a ~20 bp repeat region on its 3’ end. This repeat region associates with the trans-activating...is 3’ phosphorothioate linkage, where a sulfur replaces a nonbridging phosphate oxygen. Other useful backbone...being studied. Check out Addgene’s validated gRNA repository to find pre-tested gRNA sequences for your next...Addgene's CRISPR resources Request ready-to-use viral preps of select CRISPR lentiviral plasmids Additional...

  18. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...modification with Homology Directed Repair (HDR), CRISPR activation or repression, transgenic mouse line creation... which means they are non-infectious and non-replicative. Nanoblades are produced by transfecting the ...compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades CRISPR-based HDR allows for...transduction efficiency by neutralizing the charge repulsion between virions and the cell surface, and also...

  19. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...Associate Professor at Boston College. National reports stress the importance of providing authentic research...overexpression vector. This plasmid contains origins of replication and selectable markers that allow students to...functional MET genes to grow.) Students then use replica plating to determine if expression of MET/Met genes...complementation should only occur when galactose replaces glucose as the carbon source. Student results .... cerevisiae met deficiencies (manuscript in preparation), indicating that Met protein functions have ...

  20. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    ...including a handful of plasmids that have been in the repository for many years, has now grown into a collection...neutralization assays, part of a study that examined the repertoire of antibodies from 19 COVID-19 convalescent individuals...tools based on Cas12a and Cas13  which could be repurposed for SARS-CoV-2 RNA detection. Early in the pandemic...above, some of these plasmids have been in the repository for many years while many were developed during...Harrison SC, Wesemann DR (2021) Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike ...
Showing: 461 - 480 of 956 results