...elements, but among the most important are the inverted terminal repeats (ITRs). ITRs are the elements...2020). Adeno-Associated Virus Serotype-Specific Inverted Terminal Repeat Sequence Role in Vector Transgene...Merten, O.-W., & Galibert, L. (2017). Impact of Inverted Terminal Repeat Integrity on rAAV8 Production ..., & Logan, G. J. (2019). A User’s Guide to the Inverted Terminal Repeats of Adeno-Associated Virus. Hum...2017). Deletion of the B-B’ and C-C’ regions of inverted terminal repeats reduces rAAV productivity but...

...to access, which often forces scientists to use invasive measures to perform in vivo optogenetic experiments... channelrhodopsin-based optogenetics minimally-invasive, Frances Arnold’s lab and Viviana Gradinaru’s ...have photocurrent properties best suited for less-invasive optogenetics experiments. The sequences in this...paired with a systemic AAV allow for minimally-invasive optogenetics While ChRgers performed well in a... see if they were suited for in vivo minimally invasive optogenetics. Both experiments delivered ChRgers... In the first experiment, ChRger2 was non-invasively delivered to the dopamine producing neurons in..., ChRger2 was delivered and activated with non-invasive methods to the neurons in the right secondary ...

...we introduce you to the Journal of Emerging Investigators, an open-access journal that enables high school... the interview with the Journal of Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction...Olivia Ho-Shing, and the Journal of Emerging Investigators. Chapter 2 - Getting high school students and...achieving it. Chapter 5 - JEI's future - getting more involved in the classroom by building curricula and developing...

...reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of the DNA ...recombinase, which is named after its ability to invert or ‘flip DNA’, is derived from the yeast S. cerevisiae...Recombination between these sites can lead to excision, inversion, and translocation of DNA in a similar fashion... Mutated FRT and loxP sites in Flex Vectors – inversion only once All recombination events mediated by...target sites have been engineered to avoid this re-inversion issue. It became possible to design FLEx vectors...vectors (Flip-excision vectors) that only allow one inversion event when it was discovered that it is not the... FLEx vectors, the DNA sequence that should be inverted is flanked by two pairs of target sites that differ...

...clues about the types of neurons involved or whether their involvement is activating, inhibiting, or modulating... another. In addition to the neuronal networks involved in directly controlling muscle activity and coordinating... reflect motivational and other aspects may be involved. For example, an animal moving around to explore...norepinephrine to promote vigilance. A recent study investigated the injection of AAV8 particles expressing an...addiction and craving (‘seeking drugs’).  What’s involved in locomotor sensitization? Previous research ...circuit, and the medial prefrontal cortex (mPFC) are involved. Neurons from the mPFC project to the NAc, where....   A study from 2014 (Howerton at al. 2014) investigated the connection between neuroinflammation and...

...following strand invasion and D loop formation, the newly synthesized portion of the invasive strand is displaced...processed end of the non-invading strand at the other DSB end. The 3’ end of the non-invasive strand is elongated... one invading strand at a DSB that can be used for repair (a one-ended DSB). This single invasive strand...the canonical homology-independent pathway as it involves the direct ligation of sticky or blunt ends and...both a substrate for proteins required for strand invasion and a primer for DNA repair synthesis. The ssDNA...DSBR) In the classical DSBR pathway, the 3’ ends invade an intact homologous template to serve as a primer...branched structures that form when elongation of the invasive strand “captures” and synthesizes DNA from the...

...Testing AAV inventory for the entire collection! We are expanding our entire control AAV inventory to give...addition to our inventory of Cre-dependent GABA sensors. See all our GABA sensor AAV inventory here. pAAV.hSynap.iGABASnFR...service in 2016, we’ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of...few months. You can also browse our entire AAV inventory. Our new AAVs include: EGFP-expressing AAV for...Encoded Calcium Sensors). We are expanding the inventory of these vectors to more serotypes. We currently... GCaMP7f and GCaMP7s. See all our biosensors inventory, and check out our latest vectors below: pGP-...growing field that we aim to support by growing our inventory of recombinases and recombinase-dependent vectors...

...service in 2016, we’ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of...few months. You can also browse our entire AAV inventory. Our new AAVs include: Control AAVs for serotype...always excited to add new control AAVs to our inventory since we know that scientists need these tools...popular control AAV encoding GFP. Check out our inventory of serotype testing AAV New control AAVs Plasmid...Dana et al., 2018). See our full biosensor AAV inventory New glutamate sensors Plasmid Serotype Name.../ Chemogenetics AAV We have also expanded our inventory of chemogenetic AAV for activating and inhibiting...be activated with FDA-approved drugs. See our inventory of DREADDs / Chemogenetics AAVs New DREADDs ...

...gained invaluable experience and even after 5 years, was still improving my skills. My involvement with ...leadership training began with the NPA where I gained invaluable experience working on and later chairing the ...already have some leadership experience). I was involved with postdoctoral issues first at Brigham and ...many people at that conference, I later became involved with the NPA beyond the meeting. I first joined...where each Committee Chair and Vice Chairs were invited to spend a weekend together, funded by supporters...spoke about my work with the NPA, I was ultimately invited to join the Advocacy and Outreach Committee within... the Committee and after a year of my active involvement, I am now Chair-Elect of that Committee. Being...

...enzymes and these different Dicer-like enzymes are involved in the biogenesis of different small RNAs (Figure...Typically, the desired target of RNAi is expressed as inverted repeats separated by a loop region that facilitates..., gene knockout strategies cannot be used to investigate essential genes. Historically, gene silencing...present at the first stage of an RNAi-based investigation. Creation of the RNAi trigger requires a few...hairpin RNA The easiest approach to induce RNAi involves the expression of long hairpin RNA (hpRNA). This...of ~400 bp complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple...efficacy, thereby requiring a substantial time investment. Once the target mRNA is cleaved, the cleaved...

...of the pathways involved in the development of cancer. With hundreds of genes involved in the Ras pathway... is circumvented because of the specific genes involved. Downregulation of tumor suppressors and upregulation...on picking apart the complex cellular pathways involved in the development of different cancers and identifying...identifying the roles of key proteins involved, with the goal of being able to discriminately and effectively...pathway alone and thousands of investigators studying them, how do we ensure that the research community...Cancer Research Plasmids and Resources page, investigators may select a pathway and identify any genes ... click on a gene of interest will direct the investigator to a list of all available Addgene plasmids ...

...give you an idea of what a career in this field involves, and why you might enjoy doing it. I’ll also talk...to tackle, depending on the part of the company involved: Research & Development (R&D) teams at pharma... such teams to help make decisions about R&D investments in either new internal efforts, or through acquisitions...kinds of projects. Almost all of my projects, have involved enriching conversations with R&D, Medical Affairs...examples, and healthcare consulting firms can be involved in many similar activities depending on the expertise... Such a decision determines whether the client invests substantial monetary and human resources towards...like that would allow the client to prioritize investments over the next 3, 5, or 10 years. Complicating...

...are 11 different genes cloned into plasmids to investigate for each of these planarian species. These plasmids...time or after genetic manipulation. The technique involves using synthetically made RNA (riboprobes) with... the riboprobes are created, the WISH protocol involves preparation of the tissue sample, several washes...complex are added to the tissue. The last step involves a chemical reaction to stain the cells that contain...wildtype locomotion.   Create your own plasmids to investigate other genes in planarians! For those who are ... performing their own molecular cloning and investigating genes not in this collection, they may clone...stowers.org and if you make new plasmids for investigating different planarian genes, we encourage you ...

...work in isolation, and investigating gene networks and complex diseases that involve dysfunction across multiple...and introduce functional sequences—as is being investigated with dystrophin/Duchenne muscular dystrophy ...complexes. This enables researchers to study complexes involved in gene regulation. By generating Casilio modules...Pumilio binding sites, it will be possible to investigate the effects of inclusion or exclusion of specified...researchers in a variety of different fields and invite “developers” to build upon the Casilio platform... on engineeringbetter, more powerful ways to investigate the genome, epigenome, andtranscriptome. You ...

...available to counteract the toxin. If, however, an investigator were to successfully clone an insert into the...the insert. Gateway® technology (developed by InvitrogenTM) is essentially a more modern version of this...insertion will completely replace ccdB with the investigator’s insert of interest. Hence correct clones are... behind the “swapping out” of ccdB with the investigator’s gene of interest.   CcdB resistant E. ...CcdB-resistant strain is ccdB Survival™ from InvitrogenTM. Using either DB3.1 or ccdB SurvivalTM, one ...a variety of useful resources and protocols! We invite feedback from all members of the research community...M. “Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes.” ...

...flanking loxP sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability...sites (floxed) in opposing orientations, a SSR will invert the DNA sequence between the sites. If a DNA sequence.... Regardless, the first recombination step will invert the intervening DNA fragment using either loxP ...the additional loxP and lox511 sites after the inverted GeneR sequence in the opposing orientation, you...presence of Cre. In this example, Cre would first invert the reporter and GeneR fragment. Then the second...Sometimes also referred to as DiO (Double-floxed inverse Orientation), FLEx systems are generally considered...meaning that your gene of interest starts in the inverse/antisense "off" position and is flipped to the ...

...few inventory items offered domestically, but by the end of 2016, we expanded our viral inventory to 25... lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide...pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost entirely of viruses expressing... Carolina, Chapel Hill. These can be used to investigate the effects of activating particular signaling...neurons drives feeding behavior in mice. J Clin Invest. 2011 Apr;121(4):1424-8. PubMed PMID: 21364278....

...targeting each of the 1000 genes you’d like to investigate in your next CRISPR screen. Luckily, the Green... unbiased discovery. With these screens, the investigator generates a cell population where all genes ...consequence, which genes are good candidates for being involved in the studied phenotype. For robustness of a ... generated knockout mice for (all of the genes involved as I recall). The lab then used these knockout...sequenced to identify which signaling molecules were involved in the phenotype. Surface proteins  My group works... These types of interactions to a large extent involve receptors expressed on the cell surface. Thus, ...proteins to determine which contact points are involved in a studied cellular behavior. This approach ...

...Additionally, you are involved in multiple professional scientific societies. Why is being involved in the scientific...biology and hope to develop more in the future. Involvement in the Scientific Community Addgene: In 2012 ...ourselves without Addgene. So, when I received an invitation to join the Advisory Board of Addgene, I accepted...also beneficial to my research. I believe that involvement in the scientific community is a kind of duty...example. However, at the same time, it's fun to be involved in these professional activities because I can...

...symptoms of a negotiation, especially when money is involved. However, it doesn’t have to be this way, if you...financial values. All too often, we see early investigators making unnecessary short term sacrifices that...their goals, rather than focusing on long term investments. For example, purchasing “used” equipment can...a win-win decision An institute/department is investing in you, and is counting on your research success...solutions to reach your goals. For example, a junior investigator turned down a faculty position because none ...opinions on creating and negotiating budgets and invite you to leave a comment below Happy Sciencing, ...