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We narrowed to 790 results for: Cre

Showing: 151 - 160 of 790 results

  1. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...in therapeutics, as it can increase gene expression at certain loci to create changes in phenotypes. For... storm. While Cas9 has been most widely used to create specific edits in DNA, there has also been significant...inducible upon environmental triggers. This is done by creating a split dCas9 protein that can reassemble when...the best uses for Cas9 activators is in genetic screening. gRNAs targeting every gene in the human genome...interactions. Additionally, large scale CRISPR-based screens can be used to gain insight into protein structure...focused advancing scientific discovery through the creation of innovative technologies. To date his group ...of cancer therapeutic targets using CRISPR–Cas9 screens. Nature 568:511–516 . https://doi.org/10.1038/s41586...

  2. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    Published
    March 15, 2018, 12:59 p.m.
    ...nuclease domains, researchers created the CRISPR nickase. Nickases create a single-strand rather than a...generated by the nickases; D10A creates 5’ overhangs and H840A creates 3’ overhangs in a PAM-out design...Conversely, the H840A mutation in the HNH domain creates a non-target strand-cleaving nickase. Instead of...strands bluntly with WT Cas9 and one gRNA, you can create a staggered cut using a Cas9 nickase and two gRNAs...other? The gRNAs must target different strands to create a DSB, but this can be accomplished with either...an individual gRNA with WT Cas9, repair levels decrease rapidly 10 bp from the cut site. So if you can... you can’t obtain high HDR efficiency. Nickases create a staggered cut - could this system mediate repair...

  3. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    Published
    Jan. 22, 2019, 2:41 p.m.
    ...autophagosomes with lysosomes leads to an increase in yellow puncta but a decrease in red puncta. Find the mRFP-...RFP ratio: a decrease in GFP fluorescence relative to RFP fluorescence suggests an increase in autophagic...flux, while an increase in GFP fluorescence relative to RFP fluorescence suggests a decrease in autophagic...yeast or the lysosome in mammalian cells. This decrease in pH quenches SEP but not DsRed, leading to emission...Therefore, the formation of autophagosomes leads to an increase of yellow puncta (GFF positive/RFP positive), ...fusion of an autophagosome with a lysosome. An increase in autophagy leads to more yellow and red puncta...

  4. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ... rounds of screening. Created with Biorender.com.   Some phage display libraries screen antibody genes...serine with dispersed hydrophilic residues for increased solubility (Monnier et al., 2013). The linker ...expression systems. scFvs are also small enough to be screened for with in vitro display methods such as phage...applications where longer retention times often increase therapeutic efficacy (Ahmed et al., 2012). How...and VH sequences. Phage display also allows for screening scFv generated from the B cell antibody genes ...the ways antibodies are. scFvs are even used to screen for new antibodies. To do this, antibody variable...variable domain sequences are expressed as scFvs and screened with in vitro assays to select for strong binders...

  5. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ... are isolated from the resulting colonies, and screened for the desired modification. Finally, the positive...final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of interest...multimerization (purple) which is evident by an increase in the size of the lower band (portion of the ...mutation). Similar results can be obtained with PCR screening using the primers specified by the arrowheads ... vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward and reverse...Template A high purity plasmid prep significantly increases the success rate of site directed mutagenesis....secondary structures of the DNA template, and may also decrease the annealing temp of the primers. Because you...

  6. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...14 and 21 to create GEmCherry2. The Denham lab made several other small changes creating 2 other versions...Cas9 activity assays. Recently scientists have created CRISPR fluorescent reporter assays to quantify ...CRISPR/Cas-stimulated HDR activity) The Madison lab created SRIRACCHA in 2017 to enrich and identify desired...H2B-GFP. This visual read out allows for easy screening of different gRNAs by simply changing the gRNA...plasmid that contains Cas9. The Madison lab also created an inducible SRIRACCHA system (iSRIRACCHA) where... H2B-GFP expressing cells exhibited a 2-4 fold increase in the number of endogenous indel edits when compared... sites and activity (Knudsen et al., 2018).  To create GEmCherry2, the Denham lab first incorporated an...

  7. Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages

    Type
    Blog Post
    Published
    Jan. 11, 2017, 3:30 p.m.
    ... the injunction. While much of this decrease is due to an increase in the number of universities’ whose...postdocs who were expecting a salary increase have had that increase cancelled. Rounding out the remainder...payment for working more than 40 hours per week increasing from $23,660 to $47,476 per year. This post discusses... of postdocs who weren’t going to get a salary increase prior to the injunction still won’t after the ...issues such as childcare using money from their increased salaries. In addition, salary uncertainty may ... costs of living would get much better salary increases that those in less densely populated areas. This...Gary's Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating for Science...

  8. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ... of interest. Although efficiency may decrease with an increased number of fragments, or the ligation ... cleave outside of their recognition sequence, creating four base flanking overhangs. Since these overhangs... fragments, these problems can be overcome by screening a higher number of potential clones. Golden Gate...the ordered assembly of multiple DNA fragments to create TAL effector nucleases. These plasmids were designed...This cloning strategy not only makes it easy to create a single gRNA-expressing plasmid, but it can also...fragment: PCR-based amplification can be used to create silent point mutations at internal recognition ...Whether you are using the Golden Gate method to create CRISPR/Cas9 constructs, assemble standard plasmids...

  9. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...challenging for amateur scientists. Planarians, wormy creatures commonly found in freshwater ponds, provide a ...promoters to allow for bidirectional transcription to create dsRNA. Students can produce DNA templates via PCR... PCR off of the same plasmid and use these to create riboprobes for in situ hybridization experiments.... as how to perform worm amputations. They also created an online companion resource called “cutting class...within an embryo or tissue. Once the riboprobes are created, the WISH protocol involves preparation of the ...as a result, these cells turn blue. Students can create synthetic riboprobes off of linearized DNA templates...from their organism of interest, use the RNA to create a complementary DNA (cDNA), and then use PCR and...

  10. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...protein in your hand? At 3D Molecular Designs, we create physical models of proteins using 3D printing technology... real”.  We sometimes forget that the models we create are around 30 million times larger than the real...were soon developed that allowed researchers to create stunning, interactive images of their proteins....physical models of protein structure? We have been creating physical models of proteins for researchers for...educators and students use molecular models?   We created the Center for BioMolecular Modeling (CBM) in 1999... 1999 with the expressed purpose of creating physical models of proteins and other molecular structures...researchers to learn about their research while creating physical models of proteins that are being investigated...
Showing: 151 - 160 of 790 results