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We narrowed to 751 results for: Nes;

Showing: 81 - 100 of 751 results

  1. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM...While editing single genes is very useful, sometimes, you need to modify multiple genes to generate a cellular...adenine base editors with improved editing in cell lines Nicole Gaudelli, the Associate Director and Head...ABE8 constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The dC9-ABE8.x-m/d constructs...adenine base editors The most commonly used S. pyogenes Cas9 requires the target site in the genomic DNA...(“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites that...In addition to stem cells the authors targeted 6 genes in primary human T cells using 8 selected ABE8s ...

  2. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...carrying genes for bacterial toxins like colicin typically also carry the necessary resistance genes to help...cell to another). Bacteria need special fertility genes to be capable of conjugation, and these are encoded...organisms to thrive in competitive environments. The genes encoding these functions form the basis of many ... Resistance (R) plasmids: microbial defense Genes allowing bacteria to survive toxins are the clearest...Nevertheless, the transfer of plasmids carrying these genes between bacteria can also have serious implications...: microbial infection Virulence plasmids encode genes that allow the host to infect other organisms. If...difference is virulence plasmids! This category includes genes that help bacteria invade hosts, including attaching...

  3. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ...from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state... we can see why: two arginines make base-specific hydrogen bonds to the guanines, but the N (a thymine...    Figure 3: Crystal structure of S. pyogenes Cas9 bound to guide RNA (PDB ID 4ZT0), shown opaque...the protein, spanning both lobes. Stem loop 1, nestled between the REC and CTD domains, is required for...protein portion of the CTD, ready to check for PAM sequencines. This helps explain why Cas9 can’t locate PAM...sequence isn’t in place yet.     Figure 4: S. pyogenes Cas9 in the apo state (PDB ID 4CMP, top) compared... residues in the CTD can get cozy with DNA. S. pyogenes Cas9 recognizes the PAM sequence 5’-NGG-3’, where...

  4. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...This post was updated on Aug 20, 2020. What genes are important in your phenotype of interest? Many scientists... cause is not entirely known. Identifying which genes are important for a phenotype can lead to a wealth...experiments investigating the role of individual genes or entire pathways in a particular disease process... can also be used to activate or repress target genes without permanently modifying the genome. If you'd... is to use Cas9 and a pool of gRNAs to identify genes that are essential for a given phenotype. Image ...screen a population of mutant cells to identify genes involved in a particular phenotype. CRISPR can be...created to knock out, activate or repress target genes by combining a gRNA library with the aforementioned...

  5. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    Published
    July 20, 2017, 2:30 p.m.
    ...with other genes. He will use fluorescent proteins to monitor the localization of these genes, CRISPR to...to identify candidate genes as drivers of peritoneal mesothelioma tumorigenesis. They plan to use CRISPR...derived peritoneal mesothelioma cells lines and assess their tumorigenesis. These experiments will serve to...validate these candidate genes and allow Nabavi and colleagues to prioritize which genes they will use to develop... Di Schiavi will be able to better choose which genes and pathways to target in future small molecule ...molecules targeting SMN1 and its partners in mouse cell lines. Dr. Di Schiavi hopes that these studies will accelerate... research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic review of ...

  6. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    Published
    Nov. 25, 2013, 10:12 p.m.
    ...interspaced palindromic repeats) Cas (CRISPR associated genes) system. This popular genome engineering tool offers... be incorporated into the bacterial genome. Cas genes, which code for polymerases, nucleases, and helicases...adaptive immune system that utilized various cas genes not only to store a record of invading phages but...CRISPR/Cas is the system that scientists have harnessed for genome engineering. The diversity of the CRISPR... CRISPR/Cas systems provides powerful lines of defense against invading phages. Rising to the challenge...challenge - Evolution of anti-CRISPR genes in phages The CRISPR/Cas adaptive immune system seems like the winning...nucleases from re-identifying it (3). Surprisingly, few genes have been identified that neutralize CRISPR/Cas....

  7. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...of a gold standard set of genes (1,580 essential and 927 non-essential genes). The Root and Doench lab...viability. In addition, high copy number genes may mimic viability genes as they are subject to an increase ...help reveal the function of lowly expressed genes or genes that are more drastically affected by gene ...overexpression library identified several overlapping genes, most genes were unique to the individual libraries. These...be used to knock out, inhibit or activate target genes by combining specific sgRNAs with Cas9 or Cas9 derivatives... CRISPR libraries in the depletion of essential genes. In the subsampling analysis of the sgRNAs, Brunello...discriminate between non-essential and essential genes similarly to the Brunello knock-out library while...

  8. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...that this system uses plasmids where the genes (either viral genes or minigenome) are under control of a ...transcription (6). The EBOV genome contains seven viral genes, each flanked by a transcription start signal sequence...replication and transcription (Figure 1). All viral genes are removed and replaced by a single (non-viral)...replicates the viral genome and transcribes viral genes. Four viral proteins are required for viral genome...When used to study minigenomes, these four viral genes are each encoded on separate plasmids and are co-transfected...alongside a plasmid encoding the minigenome. These genes and the minigenome are transcribed (and the viral...is technically possible to encode all four viral genes in the same plasmid (as opposed to four separate...

  9. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions are rescued by helper plasmids...will be expressed, since the early genes are absent, and the late genes (and their structural proteins, ...the world, hypotheses began to develop that this illness was caused by either SARS-CoV-2, Adenovirus type... rarely cause severe disease in humans (and no vaccines are available on the commercial market). Although.... Its genome consists of “early” (E) and “late” genes, and the transition between early and late infection...replication (Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole...plasmids used during production and/or by genes present in the HEK293T cell line used (which have fragments...

  10. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...harmful genes into a transfer vector. Lentiviral particles expressing proto-oncogenes or oncogenes for instance...DNA efficiently into dividing immortalized cells lines but reach their limits when it comes to transferring...packaging plasmid containing all the structural genes necessary for the production of a viral particle...the packaging plasmids lacked the psi signal, the genes encoding the structural proteins could not be integrated...plasmid), 2) the HIV-1 genome containing all viral genes with the exception of the env gene (packaging plasmid...taken especially if you are working with proto-oncogenes or with human samples that have not been tested...generation because this vector requires only three HIV-1 genes (gag, pol, and rev) for production. Third generation...

  11. Advice for Moving Into Sales After Your Science Postdoc

    Type
    Blog Post
    Published
    Aug. 21, 2014, 3:15 p.m.
    ...studying cell division. However, after 6 years in the business of western blots and cloning, I decided to pack...with little emphasis placed on the marketing or business background of the applicant. However, although...push not only my academic background but also my business acumen and sales experience. Most scientists think... I tore up my “Scientific CV” and wrote a more business friendly CV that would focus on my achievements...conferences I presented at and placed them under headlines entitled “Presentation Experience”, “Career Achievements... profile, matched it with my CV and added more business-like elements. At this point you may be asking... the focus of your CV from  science-focused to business-focused?" The reason is that I knew that the mind-set...

  12. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...CRISPR/Cas9 for gene disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag...expression levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also offered clearer...development. Representative images of single focal planes through the middle of each embryo are shown, comparing...elegans: mScarlet3 for applications needing intense brightness (such as low-expression proteins of interest)...Here, the researchers were able to target multiple genes in M. abscessus either simultaneously or stepwise...e0033523. https://doi.org/10.1128/jb.00335-23.  Bioluminescent reporting of mechanical forces with PILATeS...expressed separately) assemble and catalyze the luminescence reaction (Figure 5). The researchers used the...

  13. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...plasmids containing these genes. As with the PIs above, many of these genes were discussed at the 2018...context of chromatin?” Though it’s true that adding histones and chromatin architecture to every diagram portraying...off point. The following lists of PIs, articles, genes, and blog posts are not comprehensive, so just because...Zhang Leonard Zon  You!     Chromatin-related genes with plasmids available from Addgene The links below...Keystone meeting for Chromatin. Note that histone genes frequently have many different variant names, not...our website. In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1 CREBBP... RBX1 STAG2 Xist Your favorite gene! SWI/SNF Genes:       BRG1(SMARCA4) BAF170 (SMARCC2) ARID1A...

  14. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ...tested in different plasmid backbones that varied in size or carried different genes. In these cases, gene ...three genes whose proteins are needed sequentially. When Segall-Shapiro et al. placed the genes under ...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells... fluctuations due to copy number by introducing genes of interest into the chromosome. But this has its...replication more than once per cell division and thus, genes closer to the origin of replication will be overrepresented...initiated before the cell has divided. This means that genes located near the origin will be over-represented...stabilized promoters were equivalent whether the genes were expressed from the genome or a plasmid. Thus...

  15. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...nitrocellulose membranes and exposed to IPTG in order to induce protein expression. Cells on the membranes will ...high salt.  To probe membranes with target bacterial cells, wash your membranes with PBST to remove salt...done, this technique can also help pinpoint the genes within a sequenced genome that encode for functional... vector. In order to reduce the chances of some clones being outcompeted by others in liquid culture, ...When your protein induction is done, peel the membranes off the plates and place each onto a lysis solution-soaked... gene encodes that protein! First, block your membranes with BSA or skim milk as you would for a Western...to remove cell debris. Then, UV-irradiate the membranes for 15 min to kill any remaining live E. coli....

  16. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...cell lines? One of the major uses of the piggyBac™ system is to create stable transgenic cell lines. The...method of making stable cell lines is to transfect, select, and isolate clones. This procedure is cumbersome...expression of certain genes, such as those that have negative impacts on fitness. One consideration when...including making transgenic cell lines and genetically-modified cell lines. Furthermore, the use of piggyBac...be used for numerous applications, including transgenesis, gene-trap screens, and gene editing. How does...genetic manipulations in animal models or cells lines, including stem cells, without leaving behind any... somewhere between one hundred and one thousand clones to find a cell line with the edit of interest. ...

  17. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...different Dicer-like enzymes are involved in the biogenesis of different small RNAs (Figure 1). These siRNAs...elicits immunogenic reactions that complicate the effectiveness and interpretation of RNAi. Fortunately, researchers...incorrectly relegate RNAi to history. The straightforwardness of RNAi is unmatched—unlike CRISPR experiments...the level of DNA, thereby limiting CRISPR’s effectiveness at high copy numbers and genomic loci with low...strategies cannot be used to investigate essential genes. Historically, gene silencing methods were thought...equivalently precise, but each method recovered different genes known to be required for viability (Morgens et al...list emerge that accurately reflected the scope of genes known to be essential for cell growth. Together,...

  18. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...predicted genes have unidentified roles as well (Vanni, et al). Is it true that this many genes are truly...biologically relevant situations. Tissue-specific genes – the genes of interest may not be expressed in the tissue... be observed. Functionally redundant genes – multiple genes which function in overlapping roles can...Around 20% of human genes have no known function or a poorly defined function (Wood, et al). In the microbial...line you are working with. Development-specific genes – if a gene functions at multiple stages in development...the organism you are working in. The spacing of genes and whether or not it’s ok to disrupt endogenous...construct to generate a collection of monitored genes and mutants - viral integration, transposon, and...

  19. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...permissive cell lines such as HEK293, H0 or A549 and assume that the results obtained in these lines can be translated...laboratory tool often employed to establish cell lines that stably express a gene of interest. While the...lead to low titers and poor transductions. Cell lines should be routinely subcultured at densities that... cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting ...personnel. If you are unsure of the background of the lines you are using, it is best to start from scratch.... a variety of sources such as serum, other cell lines, or infected personnel and can persist undetected...mycoplasma detection kits that use detector cell lines, ELISA, fluorometry, or PCR-based approaches to ...

  20. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...viscosus, Aerobacter aerogenes, Aeromonas hydrophila, Agrobacterium tumefaciens, Alcaligenes eutrophus, Azotobacter...salmonicida, Agrobacterium tumefaciens, A. rhizogenes, Alcaligenes eutrophus, Enterobacter sp., Erwinia amylovora...plasmids in nature, and provide tips on how we can harness those features to create broad host range recombinant...range and can replicate and stably maintain the genes they carry among organisms belonging to many different...processes, and can be used by scientists across disciplines.  What confers broad host range in plasmids? ...that the plasmid is conjugative and carries the genes necessary for transfer initiation between bacterial...Acinetobacter spp., Aeromonas spp., Agrobacterium spp., Alcaligenes spp., Aliivibrio salmonicida, Anabaena spp., ...
Showing: 81 - 100 of 751 results