...multiplexed elements. The team designed sgRNAs that include an internal barcode and separated multiple sgRNAs...was first described in Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase...coding sequences for a range of applications, including CRISPR activation by dCas9-VPR, prime editing,...REVeRT in various contexts in vitro and in vivo, including in human retinal organoids and live mice. Importantly..., Parkinson’s, ALS, and Huntington’s Disease, including plasmids, antibodies, viral vector preps, CRISPR...

...an interview next week and it includes a chalk talk. What should I include in my chalk talk?" So what exactly...experiments or a laundry list of molecules to screen has unclear utility. You cannot predict what will happen. Without...proposed approach to resolve the knowledge gap? This includes the overview and the individual aims. Describe...Howard Hughes Medical Institute. His interests include the quality control of secretory proteins in the...

...interactions. For difficult to release proteins, including those in the nucleus, more stringent buffers may... may be needed. Since many factors, including salt concentration, ionic strength and pH, affect binding...may degrade the target protein. To prevent this, include protease and phosphatase inhibitors in the lysis...Controls As with any experiment, it is critical to include proper positive and negative controls for an IP...sample that does not. Common negative controls include knockout cell lines or tissues that do not express...

... the 1930’s in a range of biological studies, including investigations into environmental pollutants and...organism with a clearly established body plan including head, eyes, and tail. Compared to the opaque mouse...process. Additional zebrafish applications and conclusions Zebrafish are also popular outside of developmental...not ideal for every application. Disadvantages include their duplicated (i.e., redundant) genome, their... these caveats, several features of zebrafish—including the available genetic resources and transparency...

... activation (CRISPRa) tools for neuroscience, including transgenic mice, neuron-optimized viral vectors... express Cre under neuron-specific promoters, including calmodulin-dependent kinase II alpha (aCaMKII;... in primary rat neurons over other promoters, including ubiquitous ones such as EF-1α and CAG. ... the differentiation of other CNS cell types, including microglia, astrocytes, and oligodendrocytes, provided...applications for which CRISPRa is well-suited, including studying mRNA dosing effects, cellular reprogramming...

...systems and creating an all-synthetic vector that included only the elements needed for function. The lab...channelrhodopsins for use as optogenetics tools. These tools include a pair of channelrhodopsins, Chronos and Chrimson...CsChR, and CoChR have been deposited to Addgene, including lentiviral and AAV expression vectors for certain... for the Rinehart phosphoprotein system, which includes detailed protocols provided by the Rinehart lab...repressor plasmids from Scot Wolfe's lab. These include Tet-inducible CRISPR activators and repressor plasmids... contain a variety of N- and C-terminal tags (including His, S-tag, OneStrep, CBP, Trx, GST, Halo, MBP...transgenesis. TheGreenGate kit available at Addgene includes six individual types of pre-cloned insert modules...

...genes that regulate a wide variety of phenotypes, including but not limited to cell growth and drug resistance...activate Ras are found in one third of all cancers, including a high percentage (up to 95%) of pancreatic cancers...crowd-sourced map of the RAS signaling pathway and includes 180 unique transcripts. Each gene was chosen to...provided in both open (no stop codon) and closed (including a stop codon) formats. The collection is compatible...collection of over 30 promoters and 140 genes, including constructs with unique tissue-specific promoters...key host for the production of their products, including fermented beer and wine, fuels, pharmaceutical...individual tagging event. Commonly used sequences, including fluorescent tags, a floxed Cbr-unc-119 selectable...

...editing to change the genetic code of plants, including the development of crops with longer shelf life...to engineer the genome of many plant species, including commonly used model organisms like Arabidopsis... Medicago truncatula and several crop species including potato, corn, tomato, wheat, mushroom, and rice... used RNA pol II promoters for Cas9 expression include the ubiquitously expressing cauliflower mosaic ...toolkits for creating plant expression plasmids, including plant CRISPR plasmids from the Yiping Qi lab and...

...that there are many companies and organizations (including Addgene), that need people who are data oriented...academic environment. Some great tips from this panel included:  Write Often Whether you take on opportunities...publishing process. The diversity panel - Equity and inclusion The final and possibly the most important panel...show that people from all backgrounds are truly included as members of their research communities and are... be asked going forward is: How do we measure inclusion and equity? This is a difficult question but not...

...episodic memory, a special type of memory which includes autobiographical events (see Jin & Maren, 2015...visualization of the morphology of individual neurons, including dendritic and axonal extensions and, most importantly...review see Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal...some engineering, RABV and HSV, but also others including pseudorabies virus type 1, vesicular stomatitis... Addgene's AAV guide)  Commonly used serotypes include AAV1 – 9, with most of these serotypes capable ...future developments in the field of AAV vectors. Conclusion An ideal comprehensive toolset for neuronal tracing...

...workplace bullying? Threats to professional status – including belittling opinions, public professional humiliation...competence procedures Threats to personal standing – including undermining personal integrity, destructive innuendo..., intimidation, sexual harassment Isolation – including preventing access to opportunities, physical or... withholding necessary information Overwork – including undue pressure, impossible deadlines, unnecessary...unnecessary disruptions Destabilization – including failure to acknowledge good work, allocation of meaningless...

...technology can be used for numerous applications, including transgenesis, gene-trap screens, and gene editing...manipulations in animal models or cells lines, including stem cells, without leaving behind any accessory...piggyBac™ transposon containing a selection marker is included in a standard homology directed repair template... used for a variety of research applications, including making transgenic cell lines and genetically-modified... using a variety of gene editing technologies including Zinc Fingers, TALENs, and CRISPR-Cas9 nucleases...

...reagents for many molecular biology protocols, including vector-insert joining for recombinant plasmid ...efficiently ligate many undesirable structures, including substrates containing gaps or DNA base pair mismatches...specificity and quantitative accuracy and even include profiling single nucleotide polymorphisms (SNPs...substrate (23, 24). Thermostable DNA ligases, including Taq DNA ligase, are naturally able to discriminate...DNA ligase commercially available (see below). Conclusion It is important to note that thermostable, high...

...high as 1-3%. In light of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors...you intend to use performs in your system. This includes a thorough testing of both the serotype and promoter... is reached you may obtain a false negative or conclude the viral vector is not working.  When using a...off-target expression that can confound your results.  Conclusion Using AAV vectors in your experiments requires...system. It is very rare for a methods section to include all of the variables that need to be accounted ...

... advice for the group of juniors and seniors, including that it’s never too early to start networking!...was the favorite part of their visit for many, including Natalie Reid who said “Seeing the bacteria on ... of programming for scientists!). Careers that include both science and business or programming were new...showed some highlights of his travels from 2015, including a great European photo slideshow from our UK team...

...2015). This kit offers a broad range of parts, including copy number machinery, chromosomal integration...YTK and adds several additional capabilities, including conditional gene expression, multiplexed CRISPR...chromosomal integration and gene expression, the system includes a toolkit to express recombinant proteins within...kit plus the plant-specific parts you‘ll need, including sub-cellular localization signals, plant markers...engineering. This is a specialized system with parts including site-specific recombinases, regulatory elements...

... impact downstream outcomes of an experiment, including the accuracy and efficacy of findings. If you’...peptides and antibodies. Example applications of this include coupling to an antibody, whereby excitation of ...them under experimental conditions. Common issues include photobleaching, non-specific binding, and quenching...biotechnology. Recent examples of advancements include the Janelia Fluor(R) (JF) dyes developed by the...super-resolution applications. Other current advancements include work exploring far-red-emitting dyes in bacterial...

...selection screening. They and their colleagues including Feng Zhang used the GeCKO library to identify ...libraries developed by MIT's Tim Wang and colleagues, including David Sabatini and Eric Lander, comprise 73,171...different gene targets. These pooled libraries include enrichment against kinases, cell cycle proteins...designed to target a smaller set of genes and it includes more guides per gene. The Sabatini/Lander library...

...submissions via email: [email protected] Entries include accession number, species, and assigned category...profiling for human, mouse, and dog cell lines is included when available, and can be submitted via email...Depending on available information, entries may also include whether a cell line is known to be genetically ... cited. If the information seems incomplete or unclear, Bairoch follow up via email to the authors of ...

...stays on the same page. Ideally, these meetings include all collaborators, from the lab members who are...like Zoom or Google Hangouts is a great way to include researchers from different locations. Having virtual...to your collaborators. This helps everyone feel included in the project and accelerates the pace at which...and morale. Especially when your collaboration includes scientists working in different time zones, it...