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...genetic modifications into mouse embryos. But once you generate a growing colony of genetically engineered...epigenetic modifications. Special breeding cases Whether your mouse colony consists of genetically engineered...Cre-lox lines, and how to properly control for genetically engineered mice in your experiment. As you’ve...ve learned in Part 1, there are many types of genetically engineered mice: transgenic mice, knockin and...techniques are each useful for introducing one modification into the mouse genome, they are not commonly...parental mice In order to ensure that your genetic modification will be passed along to all of the offspring...getting a small piece of tissue from each mouse (typically via either a blood draw, tail clip, or toe clip...

...characters. A descriptive purpose focuses on the practical application of the plasmid and will help a scientist...species can be indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid...usually very easy — our tech transfer team will communicate with your organization’s technology transfer ...name and match the name as it appears in its publication or as it is commonly known in your lab. Plasmid...reagent/antibody, unspecified, or other. If other, indicate type in “Vector Type Other” column. For example...or room temperature. Growth Strain Please indicate which growth strain Addgene should propagate your...For plasmids that contain the R6Kγ origin of replication, choose the Pir1 strain. If your plasmid cannot...

...lentivirus, AAV does not possess the ability to replicate itself and depends on helper viruses like adenovirus...herpes simplex virus to facilitate efficient replication. The most common production system, however, ...RNA genes; and the pRepCap plasmid containing replication factors and serotype-specific capsid genes.  ... steps of AAV production, iodixanol gradient purification, and AAV titration by qPCR.   Pro tip! An optional...al., 2010). For certain in vitro and in vivo applications, you can consider using unpurified AAV from ...Benyamini et al., 2023) to save time and cost.  Purification To purify your rAAV from your supernatant, you...recommend using a Tris-based lysis buffer and sonication.  The obtained virions from cells and supernatant...

...easier of the two, but it actually requires more technical skill to place the needle in the correct vein ...institutional oversight is required. Restraint: mechanical restraint or anesthesia Location: tail vein or...you might after getting a vaccine. Restraint: mechanical restraint Location: thigh Gauge: 25–27 Volume...xenograft models. Restraint: back scruffing or mechanical restraint Location: back or hind legs Gauge: ...between the layers of the skin. While this could theoretically be performed anywhere on the body, somewhere... correct location requires a lot of skill and anatomical knowledge. Intrathecal injections are used for...stomach. This technique involves administering medication through a feeding tube with a bulb on the end...

...What’s behind these sights are an abundance of biological molecules like chromoproteins, fluorescent proteins...proteins and luminescence have been widely used in biological research for a long time, chromoproteins are ...just now being developed for a broad range of biological inquiries.  What is a chromoprotein and how ...photosynthetic activity. This still allows photosynthetically active wavelengths to reach zooxanthellae... sequencing are more definitive, but visual identification is a good place to start before taking on more...resources are limited or for bioart and teaching applications. Find a chromoprotein for your experiment! ...events: The plasmid carrying the desired genetic modification integrates into the genome (Figure 3). Here,...

...CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods...well as suitable applications. Expression System Components of System Application Mammalian expression...Select the appropriate CRISPR tools for your application: Wildtype Cas9 or Cas9 nickase are appropriate... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator...or more sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair... be constitutive or inducible. U6 promoter is typically used for gRNA. May contain reporter gene (e.g....key features of common  delivery methods below: Chemical transfection   Lipid mediated Cationic polymers...

...Lee and Pantelis Tsoulfas of the Department of Neurological Surgery at the University of Miami. The beginning...expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile tract, Cu= Cuneate...originally named orthogonal-plane fluorescence optical sectioning (OPFOS) (2). Arne Voie, David Burns ...of LSFM microscopes (3,4). After this initial application, Stelzer’s group described the single-plane or.... confocal microscopy In confocal microscopes, optical sectioning of a specimen is based on discriminating...used in LSFM. In LSFM the laser light sheet, typically 2-6  microns, illuminates only one thin plane ... milliseconds to acquire one image, thereby significantly reducing the time it takes to image through ...

... to be crazy complicated! Listen to the full podcast for more great science communication tips or listen...tons of ways you can get involved in science communication. In this second half of our conversation with...many things you can do start your own science communication student group and get more involved with your...is a graduate student group with the goal of communicating science to the general public and training the... the next generation of science communicators. In this chapter we learn about some of their events - science... peer-to-peer feedback in effective science communication. Chapter 7: The Goal of the SITN Podcast - Vini...Pint" event is the best way to get a Science Communication group started. It's as simple as getting people...

...transformation. A given insert will typically contain both a protein modification sequence and a selectable gene...nearly all domains of life repair genomic damage, specifically double strand breaks. Researchers have long ...tags have been integrated. Simply design your amplification primers with the desired targeting homology—... green, and red FPs that are codon optimized specifically for expression in yeast. These tagging vectors...This collection is ideal for tandem affinity purification of protein complexes. John Pringle and Jürg ...deposited a large collection of plasmids for genome modification in yeast developed by Dr. Pringle’s former lab...collection of plasmids for a wide variety of genome modifications in S. Pombe, including full and partial gene...

... plasmid or group of plasmids' functions and applications. We hope that these articles make it easier ...manipulating interneurons using a viral approach Genetically modified organisms like transgenic mice are important...FLIM (Fluorescence Lifetime Imaging Microscopy) applications. To demonstrate the power of mCyFP1, the Lin... cell physiology and molecular processes via genetically encoded proteins. PhoCl is a new member of this...experiment, check out our recent guest blog series: A Practical Approach to Choosing the B(right)est Fluorescent... Coselection efficiency is so high that some applications may permit working with edited “pools” of cells...showed high editing efficiencies with AsCpf1, indicating that ouabain coselection is a robust and flexible...

...backgrounds with technical expertise that ranges from astronomy to bioethics to chemical engineering and...Citizenship Initiative.    Get practical experience in science communication by writing for the Journal of...societal pressures including economic, cultural, and political. Science policy looks differently depending on... policy implementation, program management, communications, diplomacy and international relations, advocacy... p” policy.  “Big P” policy is what you might typically think of. It occurs in a government setting: drafting...STPF cohort members came to DC to do either a sabbatical year or transition away from academia as tenured...move into science policy. Read broadly about the historical roots of science policy to connect to your why...

...culprit for these unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one...Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing a lack...transcription) is demarcated by viral genome replication (Harrach, 2014). Given that the early genes ...encapsidate new virions) are not expressed. Note that replication-competent AdV vectors also exist and are used...the final topic, which is that natural AdVs, replication-competent viruses with a recombinant genome, ...basic research, like other viral vectors, though significant challenges remain (Wold & Toth, 2013).  While...Hermiston, T. W. (2003). Multigene expression from a replicating adenovirus using native viral promoters. Mol ...

...club occurs with a small technical team (single lab or team), a deep technical dive on single papers could...emerging work by collaboratively discussing recent publications in a specific field(s). Most anyone who has ...the company at large, by polling and informal communications we made a decision to try out a Bioinformatics...topic, there are a number of administrative and practical decisions to make. How often will you hold it?... the time recurrently carved out. This notice typically should include the paper, any supplemental data...repositories, or other information needed by someone to critically discuss the paper.  Before the first journal ...major findings and then focusing more on broader implications rather than diving deep into the individual ...

... general public. Chemical Society in University College Dublin, Ireland The Chemical Society was founded...way up to professors. Oxford Biological Society, England The Biological Society based at the University...idea of the Freshers Fair and some of the great biological societies that Addgene has had the opportunity...at Freshers Fairs in the past year. Cambridge Biological Society, England Since 1996, Cambridge Biosoc...Biosoc’s goal has been to promote interest in the biological sciences amongst the student community at the...sessions to help undergraduate students with their applications. The society currently has 1,283 members, who...At Addgene, we’re incredibly glad to see these biological societies engaging with young scholars. We believe...

...over a month ago I finished up my PhD in Biological and Biomedical Sciences at Harvard University and entered...talking to people about and communicating science. The path to science communication This may come as somewhat...researchers gain doing PhDs (i.e. technical writing, teaching, presenting, analytical thinking, time management... I would ultimately want to go into science communication but thought that I would be best poised to share...PhD myself. To continue to foster my science communication skills, I took on TA positions, tutored fifth...largely my work developing myself as a science communicator that got me here! Thus far being an Outreach... one of my supervisors warned me, the swift gratification of publishing posts can be addicting. Even more...

...design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were reported...enzyme can lead to off targets and result in significant, negative effects in RNA targeting experiments...originally suspected that this new Cas7-11 might be phylogenetically classified as the more complex Class I systems...non-specific activity. Its major advantage is the significantly reduced amount of off-target toxicity compared...using Prime editing alongside some creative (and significant!) protein engineering. They combined Prime editing...place, while increasing the number of potential applications. But they weren’t done engineering quite yet...switched up the serine integrase for one which can enzymatically take a genetic sequence and integrate it into...

... depends on the biological question, and zebrafish are not ideal for every application. Disadvantages ...have been used since the 1930’s in a range of biological studies, including investigations into environmental... are powerful resources for investigating any biological process. High quality genome sequence A high-...mutant, which regulates a developmental pathway implicated in congenital heart defects and cancer. These...thousands of mutations in genes affecting diverse biological processes—not just development—are available ...DNA into the genome is useful for a variety of applications. For example, specific tissues such as blood...pathways control this process. Additional zebrafish applications and conclusions Zebrafish are also popular outside...

...the Janelia Research Campus of the Howard Hughes Medical Institute. Are you currently on or planning to .... Given this large sum, one might expect that applicants would work for months to hone a chalk talk. Yet...research vision is your approach to resolving a significant knowledge gap in your field or even creating ...rejection of a model and no clear alternatives. Typically, a research vision is broken up into three aims... basis of a single grant. Either way, think realistically about the time and resources required to feasibly... aims in more depth in the remaining time. The typical chalk talk structure is: What is the big question...field, and interesting. For example, “there is a significant need for healthy livers for transplants. Growth...

..., Kyung-Mi, et al. “Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631...amplifying RNA sequences that could bind very specifically to target molecules. Using a technique called... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors... render this approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...

...formal scientific communications, they are fantastic fodder for science communication. The trick is to ... is far from the only way to communication science. Science communicators also use the hero’s journey,...this small glimpse into the world of science communication and can use what you've learned to share your...Finding Conflict in Science  In all science communication, what you are doing is telling a (nonfiction...details often aren’t included in scientific communications, but they’re firmly part of the story to the...conflict gives you an easy framework with which to communicate your science. The act of building towards your...your results right at the beginning of your communication, use conflict to get your audience invested ...