Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 902 results for: Nes

Showing: 21 - 40 of 902 results

  1. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...assembly methods. A new method, REPLACR-mutagenesis, harnesses the power of bacterial recombineering to...REPLACR-mutagenesis could save busy scientists a lot of time. Let us know how REPLACR-mutagenesis works ...Comparison of REPLACR-mutagenesis with other common methods for site directed mutagenesis   References 1. ...al. (2016). “REPLACR-mutagenesis: a one-step method for site-directed mutagenesis by recombineering.” ...Site-directed mutagenesis (SDM) is one of the key tools researchers use to prove causation in molecular... Plasmids After PCR). Current site directed mutagenesis methods Current SDM methods rely on in vitro ...structure or too much sequence similarity. REPLACR-mutagenesis: Letting bacteria do the work If you’ve conducted...

  3. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...yeast gene of interest. These fluorescently tagged genes are expressed under native conditions and allow ...looking at their performance in categories such as brightness, stability, and disruption of the tagged protein...be interested in attaching epitope tags to their genes of interest, allowing for easy capture and detection...

  4. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...effects. One-pot saturation mutagenesis The one-pot saturation mutagenesis technique described in Wrenbeck...saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How .... “Random Mutagenesis by Error-Prone PCR”. Methods in Molecular Biology In Vitro Mutagenesis Protocols...Efficient, Expansive, User-Defined Mutagenesis.” Ed. D. Dafydd Jones. PLoS ONE 7.12 (2012): e52031. PubMed...the Whitehead lab developed One pot saturation mutagenesis as a quick and easy technique that can be used...library generation prior to one pot saturation mutagenesis Several methods can be used to create mutational... shortcomings compared to one pot saturation mutagenesis. Error-prone PCR: This method amplifies the...

  5. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ...temperature ranges and make them grow better at suboptimal ones, which could allow even your regular E. coli to ...where we could incorporate cold-tolerance inducing genes within a regular mesophilic host system. This would...folding at low temperatures, since mesophilic chaperones would not function well under suboptimal conditions...engineering a plasmid which contained cold tolerant chaperones (the Cpn 10/60 isolated from Oleispira antarctica...the figure below shows, E. coli expressing the chaperones protein resulted in higher levels of growth compared...

  7. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...bound to modified proteins called histones. We can detect modified histones with “GPS for proteins,” ChIP-... of modified histones, a genomic region that has mono-methylated and acetylated histones, but not tri-...which NANOG-related genes are critical to stem cell survival. Knowing which genes influence the survival...Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome ...tri-methylated histones, functions as an enhancer. We can therefore denote enhancers as follows: (mono_methylation...combination of coiled DNA and structural proteins called histones. Chromatin folds into secondary structures (loops...

  8. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ...coming from.   Acoustic reporter genes (ARGs) Acoustic Reporter Genes (ARGs) are engineered gene clusters... reporters has its drawbacks. Acoustic reporter genes (ARGs) overcome these limitations by using gas vesicle... require injection of substrate to produce bioluminescence. Substrate pharmacokinetics and biodistribution...localization: In vivo imaging with fluorescent or bioluminescent reporters usually only detects the signal of...and high spatial resolution. By piecing together genes from gas vesicle operons from two different bacteria...their ultrasound signal. Using acoustic reporter genes for in vivo imaging Following in vitro characterization...S.P., & Shapiro, M.G. (2018). Acoustic reporter genes for noninvasive imaging of microorganisms in mammalian...

  10. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used to study...Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes that, when expressed, enable cells... vivo.  But what ultrasound lacked was reporter genes that could produce ultrasound contrast. That is,...gas vesicle-expressing species. The gas vesicle genes from Serratia sp. 39006 produced the highest levels...bARGSer expression, from left to right: colony whiteness on inducer plates, white inclusions in cells with...cell line with strong gas vesicle expression and robustness without chromatin-opening agents. It ended up...being very straightforward to engineer various cell lines from different tissues and mammalian species to ...

  11. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9" from the Journal of Visualized ...Cas9 to create genomic deletions in mammalian cell lines. Below Bauer and Canver discuss the motivations ... and colleagues using zinc finger nucleases to harness non-homologous end joining repair (NHEJ) [1]. Our...targeted deletions is to produce monoallelic deletion clones. Cells with just a single intact copy at a given...cells can be easily identified. Even in the case of clones with monoallelic deletion, the nondeleted allele...validation, and screening by PCR for biallelic deletion clones:    Like the video? The Orkin and Bauer lab's...enhancer of BCL11A subject to genetic variation determines fetal hemoglobin level. Science 2013 Oct 11;342...

  13. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    ...immortalize CDG patient-derived immune cell lines and use these lines to study how immune processes and biological...research on diseases of glycosylation, arginine mutagenesis, and neural development. Paula Videira: Congenital...studying the function of the mouse Nfia, Nfib and Nfix genes in brain development since 2000 and have shown several...

  15. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ... our paper, “Synthetic recombinase-based state machines in living cells” (1), into a more straight-forward...our engineered devices, recombinase-based state machines for your own experimental needs. For more context...timing of these factors. Recombinase-based state machines (RSMs), which take on a particular DNA-sequence... from different recombinases are overlapping or nested on a register, then the operation of one recombinase...replication), contains large serine recombinase genes downstream of inducible promoters: the BxbI gene...The paper also demonstrates that genetic parts (genes, terminators, and promoters) may be interweaved ...predictable gene regulation programs that specify which genes should be expressed or not expressed in each state...

  16. Measuring Kinase Activity at the Single-Cell Level with Kinase Translocation Reporters (KTRs)

    Type
    Blog Post
    ...localization signal (bNLS) and a nuclear export signal (NES), both which are phosphorylatable. Prior to phosphorylation... phosphorylation bNLS activity is suppressed and NES activity is enhanced, leading to cytoplasmic translocation...of the phosphorylatable residues and the bNLS and NES sequences, which have previously been characterized.... for more details on the design of the bNLS and NES sequences. Disadvantages of KTRs KTRs also have some...

  17. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ... fragmented genomes is the key to fitting large genes into AAV First, some history. In 2008, the Auricchio...mini-dystrophin.  3. Hybrid  Overview: The hybrid approach combines the overlapping and trans-splicing methods. The... A (2008) Serotype-dependent packaging of large genes in adeno-associated viral vectors results in effective...Chamberlain K, Riyad JM, Weber T (2016) Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated... Auricchio A (2013) Effective delivery of large genes to the retina by dual AAV vectors. EMBO Mol Med ...

  18. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ...repress the expression of genes primarily involved in development. These genes are known to play a role...many contexts within the fly including oogenesis and cytokinesis. Why name this protein after a vegetable..., two genes important for mesoderm differentiation Now, let's turn tinman and bagpipe, two genes important...bagpipe are both homeobox genes that are part of a genetic cascade that determines the cell fate of the dorsal...two homeo box genes that determine cell fates in the dorsal mesoderm of Drosophila." Genes & Development...they can name a gene and many of the interesting ones come from the fly community. Not surprising, this... our repository that you can use to study these genes! Groucho, a transcriptional co-repressor The first...

  19. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    ...2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes in mammalian...through a host organism, the expression of some genes can change depending on the location. Current methods...of microbes rely heavily on fluorescent protein genes. These optical reporters have limited deep-tissue...vivo using ultrasound. These ARG plasmids contain genes for “gas vesicles,” a gas-filled protein structure...demonstrates engineered gas vesicles as the first reporter genes using ultrasound and provide a major benefit to ...active miRNA’s unique to HEK293FT and HeLa cell lines. When the miRNA activity profile for a given cell...segment Understanding the connection between specific genes and an immune response has been a longstanding challenge...

  20. CRISPR History and Development for Genome Engineering

    Type
    Collection
    .... 25(10):1581-9. PMID: 26355004 Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl... anti-CRISPR genes in bacteriophages infecting Pseudomonas aeruginosa . Anti-CRISPR genes employ varied...system - utilizing various CRISPR-associated (Cas) genes to not only store a record of invading phages but...Cas proteins. Makarova et al. ’s classification defines 5 types and 16 subtypes based on shared characteristics...genomic DNA. Type II CRISPR was the first system harnessed for genome engineering, with Type V following ...Csf1) Type VI (Cas13) Fighting Back: Anti-CRISPR Genes in Phage The CRISPR adaptive immune system seems.... Pooled gRNA libraries can be used to identify genes that are important to a given phenotype. Current...
Showing: 21 - 40 of 902 results