...and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...unsure of the next steps. Harvard PI Angela DePace has put personal development at the center of her lab... and postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine...technique are beneficial for researchers at all stages. Rather than succumbing to academic inertia, it...

... and software development has caught up with it. The peer review process has been challenged with many...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable...needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve dubbed... genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research activity...significant improvement. As a result, the software piñata has been filled with slight variations on a theme without...evaluate off-targeting, but currently no software has been designed for this purpose. This second area ...findings to help everyone else. CRISPR technology has reached into so many different communities that it...

...et al., 2016; Duret et al., 2019). But the field has sometimes seen controversy, and the interactions ...yet to be demonstrated (or invented), its behavior has already been successfully reproduced by several other...tools. And, since MagLOV’s parent protein AsLOV2 has already been adapted into a variety of optogenetic...  Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet...effect (many do). Fusing EGFP to the self-labeling FlavinTag made a reliable one-component magnetoresponsive... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...

...the State Museum has been the best supporter and most productive partnership CINSO has had. Volunteer! ...provide online. Twitter is an excellent resource that has gained a lot of traction with researchers. It is ....  Try new things to further your career Take advantage of opportunities provided by your programs and...including workshops and networking events. I took advantage of as many of these resources as I could. Networking...front work was done for me. I just had to take advantage of it. Blog and tweet I started my blog, Born ... been “traditional,” I was able to fully take advantage of my graduate experience. Melanie Fox is currently...

...cleavage with some of these peptides, no consensus has been reached on which peptide works best. Likely ... is easiest to start with a plasmid that already has the multicistronic element and reporter cloned in... unique genes, you can start with a plasmid that has multiple cloning sites flanking the multicistronic...expressed from the same cassette is sometimes advantageous, particularly when only a portion of the plasmid... mRNA. However, some bicistronic vectors take advantage of an element called an Internal Ribosome Entry...bicistronic vectors; however, they do have some disadvantages. These elements are quite large (500-600 bp)...vector.3  2A Peptides To overcome some of the disadvantages of the IRES element, scientists have adapted...

...Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing field by making experimental...often under-appreciated or over-looked.  Synthego has developed a new tool called ICE (Inference of CRISPR...increased flexibility and scalability, the ICE software has two analysis formats: sample by sample analysis, ...comparable (within r2=0.96) to that of NGS data. Advantages of ICE Determine Editing Efficiency and Types...of CRISPR editing. ICE software identifies the percentage of genomes that have been successfully modified...(SpCas9). ICE Score - The editing efficiency (percentage of the pool with non-wild type sequence) as determined...size of the insertion or deletion along with the percentage of genomes that contain the indel. The discordance...

...target in cis and has no trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its...targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and variants ...scientific community. What are the features? CasPEDIA has collected relevant information on all the Cas enzymes...learn about Cas domains. CasPEDIA either directly has this information or links out to this content and...

...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...

...amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher throughput...abundance. If a scientist wants to know if an organism has a particular gene mutation, southern blotting can... DNA is complementary to the probe (for example, has the sequence of the mutated gene the researcher is..., two DNA fragments are observed because the DNA has the site where MstII cuts. In a patient with sickle...DNA fragment is observed. Today Southern blotting has been largely replaced by real-time PCR to answer ...questions (see below). Importantly, although their role has changed, restriction enzymes are certainly not dead...

...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...