...translation inhibitors cycloheximide and puromycin, Halstead et al. verified that translation is necessary ...translation is known to be important in development, and Halstead et al. used TRICK to examine translation of Drosophila...Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...

...two genes by one Gal4 driver will produce roughly half as much protein as overexpression of each gene individually...384–391 (2004). PubMed PMID: 15262411. 4. T. D. Southall, D. A. Elliott, A. H. Brand, The GAL4 System :...Marka, J. C. Kagan, Environmental Stress Causes Lethal Neuro-Trauma during Asymptomatic Viral Infections...

...about 65,000 messages and dealing with email can be half of my day sometimes.” He points out that the copious...the influence from his two PhD advisers who were “chalk-and-cheese”. While one was an established Professor....” Take home message Starting a lab can be a challenging process. Each PI emphasizes the importance of...

...longstanding challenge. Using shRNA, TALEN, or ZFN to edit immune cells poses challenges such as the inhibition... brings together the split botulinum neurotoxin halves to enable VAMP2 cleavage, and suppresses neuronal...

...components of EvolvR and its potential applications. Challenges of existing mutagenesis methods Several continuous...drawbacks with these approaches that makes it challenging for users to achieve targeted diversification...PMID: 30253789. PubMed Central PMCID: PMC6154929. Halperin, Shakked O., et al. "CRISPR-guided DNA polymerases...

...movement is that it provides interesting and unique challenges for intellectual property lawyers. While there...be, an interesting legal and professional challenge.Paratore: The promise there is the ability share the...St. Elizabeth’s Hospital and was associated with Hale & Dorr and Crowell & Morning. Karen was a founding...

...computation work comes with new sets of challenges. One challenge with computational analyses is to keep... are doing what you are doing. Another common challenge is to keep track of what works, and what does ...

...year (12% in 2018) since 2005. We’re meeting the challenges of DNA distribution to ensure scientists everywhere...happening. The Addgene team will have some fun challenges this year and I’m excited about this, but definitely...talent pipeline. Turnover at Addgene is about half of what is standard in most sectors. The Addgenies...

...quantification of enhancer activity have remained challenging due to the variable availability of enhancer ...pTALYM3, pTALYM4 Boris Greber pTAL7a, pTAL7b Michal Zurovec pBlue-TAL Nathan Lawson pJDS Series ...degredation is the speed at which it works--typical half lives of AID-tagged proteins after the introduction...studying essential proteins. Until recently, it was challenging to apply the AID-tagging system to essential ...expressing and purifying human kinases remains challenging. Small molecule kinase inhibitors usually target...

...Disease (SCD) and Transfusion-dependent beta thalassemia (TBT), both of which affect hemoglobin. Most ... enzymes target several strains of E. coli and lethally chop their genomes into many, many pieces. Locus...technology is rapidly evolving, and many of the challenges with therapies involve patient safety, effectiveness...

...experiments are either not accessible or too challenging for amateur scientists. Planarians, wormy creatures...available planarians: Girardia sp., Dugesia dorotocephala, Phagocata morgani, and Phagocata gracillis....differ. For example, Girardia sp. and Dugesia dorotocephalacan can be fed dsRNA-expressing E. coli that are...

... cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery technologies have...treatment appears to be both safe and effective. Challenges for AAV gene therapy Although AAV-based gene ...promising, scientists are working to overcome key challenges to their efficacy:  Pre-existing immunity: Many...

...infected host cell is eventually lysed. Cell lysis halts protein production, but there are non-lytic insect... however, are expensive and very technically challenging to use. Alyssa D. Cecchetelli is a Scientist... expression in Escherichia coli: advances and challenges Production of recombinant proteins in plant root...

...the tag for each insertion. Also, it remains challenging to screen for the genetically modified strains...fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP) tags, a selectable marker (floxed Cbr-unc...small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed a polymerase...

...which creates a challenge when studying them since their complete knockdown is lethal. Incomplete or titratable... to lab-adapted bacteria because it has been challenging to introduce CRISPRi machinery into diverse bacteria...

...be split into two halves, which are non-fluorescent on their own. If the two halves are in close proximity...determine the proximity of two proteins fused to the halves of the split FP. This technique is also is also...

... peripheral nervous system presents a special challenge to scientists. What works well in one cell or ...vectors in neuroscience Let’s consider my first challenge when I started working with AAV vectors. I wanted...Adeno-Associated Viral Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https:/...

...hematopoietic disorders sickle cell anemia and beta-thalassemia. Lentiviral vectors are promising agents for ...genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783-4. ...HMGA2 activation after gene therapy of human β-thalassaemia. Cavazzana-Calvo M, Payen E, Negre O, Wang G...

...gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb...transgenes might seem daunting, it’s actually a challenge that the field has overcome with split AAVs. In...WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang...

...RNA is not inherently fluorescent and can be challenging to track intracellularly, but aptamers permit... when bound to DFHBI despite being approximately half the length (Warner et al., 2014); its length reduces...next additions to aptamer soup! References Alam, Khalid K., et al. "A fluorescent split aptamer for visualizing...