...Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab has switched over to...to pTOX. However, we feel it has clear advantages, and are hopeful that continued development of allelic..., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless mutations,...isolated are those where the allelic exchange vector has undergone single crossover and integrated into the...the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange system, he is so excited...

...refined to create Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing...example, Brainbow-1.0 expresses RFP when the construct has not undergone recombination. This default state was...neuronal pThy1-Brainbow constructs, Addgene also has two Brainbow adeno-associated viral vectors (AAV)...Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as ...neuromuscular junction. In zebrafish, the method has become very useful in lineage tracing; “Zebrabow”...Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section of...AAV) available - AAV-EF1a-BbChT and AAV-EF1a-BbTagBY. These constructs contain two XFPs each due to size...

... animal moving around to explore the environment has a different motivation (and brain areas associated... certain forms of drug addiction. When an animal has been exposed to morphine or other stimulating drugs...involved in locomotor sensitization? Previous research has shown that regions such as the nucleus accumbens ...dark compartment (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors... tests.    Forced swim test The forced swim test has been used since the 1970s to test the efficacy of...rather than being out in the open (risky). Taking advantage of these characteristic traits, rodents can be...

...beginning to see Addgene as a large data set. Addgene has over 65,000 plasmids in the repository, each verified...predictors because they are just that: rare. “Nature has many genes where there are not rare codons and many...at the time, got involved. Along with Philipp Pfingstag, a visiting graduate student from TU München at...intrinsic properties (GC content or rare codon percentage) and comparative properties (percent sequence...algorithm, they found that  GC content and rare codon percentage were not good predictors. There is not much variation...  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows ...

...antibody is biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated...detecting one protein from the sample. A direct ELISA has many fewer steps than a sandwich or competitive ELISA...linear portion or the OD of your experimental samples has reached 4.0, you can resolve this issue when you ...on the Addgene blog Learn about the advantages and disadvantages of using monoclonal antibodies. Read...

...generally mediates broad cell tropism and has a high percentage of neuronal targeting after stereotaxic ... damage to the brain tissue. The injection speed has to be slow to allow the viral particles to diffuse...order to minimize the pressure into the tissue, but has to be large enough to target larger areas, typically...Alternatively, for larger brain areas, repeated staggered injections may be considered. Timing: High expression...

... and serotype, respectively. For example, AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors... components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your gene...usually quite stable and in some mammalian tissues has shown expression for several years (Wonjo et al.,...scAAV vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) ...specific cells and/or tissues is one of its biggest advantages. Specificity can be achieved using specialized...

...can also be packaged into AAV. NmeCas9 has the added advantage that it can be turned off by anti-CRISPR... 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and ...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room for additional...the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type counterpart...daunting, it’s actually a challenge that the field has overcome with split AAVs. In general split AAVs break...

...quantified by cell transduction assays. Wild-type AAV2 has been reported to have a near-perfect physical-to-...performed on all new viral vector preparations. Addgene has used this method to generally validate our viral ... Material (AAV SFM), which is an AAV sample that has been quantified by 16 labs around the world and can...Hindson et al., 2011, Hindson et al., 2013), which has been shown to be more precise and more repeatable...2011): 8604-8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL,...wild-type AAV2 stocks (Grimm et al., 1999). The percentage of genome-containing viral capsids is typically...

...measuring DA and other neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid... dLight1 dopamine sensor In 2018, Lin Tian’s lab has developed dLight1, a molecular tool to detect and...here! The GRABDA dopamine sensor Yulong Li’s lab has created the GRABDA sensor based on a similar strategy...collects the perfusion liquid for quantitation. The advantage of this method is that it can be performed in ...

...version of PubMed.  Key features This new version has an improved search algorithm which was trained on...your search terms. Key features Semantic Scholar has several ways to focus your search results. A bar ...paper’s entry, the number of citations that paper has is displayed at the top of the page, as well as the...saved to a Semantic Scholar library. You can add hashtags to library entries, which can help you organize... papers are published, similar to Facebook or Instagram. Key features Setting up a personalized feed is...

...power of science to improve the world around us has long been known; ensuring all have access to this power...information becomes easier to access by the day, it has never been more crucial to understand just how we...into a summer school course at the University of Otago’s Department of Science Communication the following... The Owheo building, home of the University of Otago's Science Communication Department The power ...Alex Gilbert is a PhD student at the University of Otago. Studying in the same Department of Science Communication...

...Mentee. A Mentor will be more effective if the Mentee has done the self-reflection necessary to identify areas... you have nothing to give, think again. Everyone has knowledge to impart. An undergraduate can mentor ...relationship, but if you don’t take this past the chatting stage you will rarely make any change in your skill sets... the helpful accountability that is one major advantage of working with a mentor.The responsibility of...graduate student – you get the idea. There are many advantages to being a Mentor: Learn other perspectives ...

...   Fig. 1: SpCas9 has sparse PAM coverage in the human genome. SpRY Cas9 has PAM sites covering the... a PAM with no sequence constraints, or at least has looser or different requirements. Luckily, there ...alternative PAM requirements (Zhao et. al, 2023). xCas9 has been shown to be less effective at NGH PAMs, though... PAM requirement, and how these proteins are advantageous in today’s genome editing landscape. The need...

..., that informs us that this antibody has specific binding and has a strong signal. We call the unique ...strong signal has a shorter exposure time than the negative control, and a weak signal has a longer exposure...rAbs are a relatively new technology that takes advantage of DNA sequencing to turn mAb clones into expression...

...easily assemble multiple DNA fragments. This system has been used to construct plasmids for use in model ...Cyanobacteria, a phylum of photosynthetic bacteria, has increasingly been recognized for its biotechnological... encoded voltage indicator Article contributed by Shreya Vedantam Genetically encoded voltage indicators...GEVIs) are proteins that detect and output the voltage changes in neural activity when paired with appropriate...appropriate fluorescent optics. Existing red fluorescent voltage indicators, however, fall short compared to GFP-based...containing a red-shifted Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin (Ace2N) with the...amino acid linker between them. The Ace-mScarlet voltage indicator works like many FRET-opsin GEVIs. With...

...industry just wasn't for me, and as has been highlighted recently it has become more difficult to attain ...interesting and luckily genetics is a sector which has been rapidly growing. These factors left me optimistic...because it is a small and dynamic company and so it has the flexibility of a ‘start-up’. I was surprised ...schedule.  I really love my highly varied role, it has allowed me to travel and to share my passion for ...everyone. My career path may seem quite unusual but it has allowed me to try out working in many different sectors... still feel now, that it is important to take advantage of the fact that science is a global language ...

...host cells immediately prior to their transduction has been shown to increase AAV transduction efficiency... such cationic reagent that is commonly used and has been shown to increase transduction efficiency by...single-use aliquots. Never use a polybrene solution that has been thawed more than once. Fibronectin is another...polybrene (e.g., hematopoietic or primary cells), and has been shown to increase transduction efficiency by...packaging cells after 1–3 days of selection. A lower percentage of surviving cells may be the result of plasmid...

...Griesbeck adds that this color change has a particular advantage for observing events taking place inside...Griesbeck of the Max Planck Institute for Neurobiology has been working on genetically encoded indicators of...Maarten Merkx of Technische Universiteit Eindhoven has similar interest in FRET-based sensor proteins, particularly...

... techniques is a speculation of yesteryear which has been realized recently. Science Fiction (SciFi) writers...even before the discovery of genes themselves.  How has scientific progress in genetic editing impacted the...technology on various non-scientific platforms, CRISPR has also transcended into the world of SciFi. This is...and most important genres of literary fiction. It has always served as a vision of the future and to make...and agriculture, its portrayal in popular fiction has been largely negative. But these negative depictions....1126/science.1258096 Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2018) Publisher... movie version (2016) stars Tom Hanks as the protagonist (Langdon). The plot involves Langdon chasing ...