We narrowed to 868 results for: REN

...deposits to Addgene.  Insomuch as different journals represent different research areas, target readers,... this story had us wondering: is there a larger trend here? Do papers associated with Addgene deposits... would also be great to know how much of that difference in citations is caused by depositing a plasmid...wouldn’t be causal. To control for a number of differences between those papers with Addgene plasmids and..., so our estimates now take into account any differences in citations due to the journal of publication... the same journals! So, is all this remaining difference caused by depositing the related plasmids with...it is still suggestive that we see such a big difference in citations after controlling for journal. And...

...  7 ways to strengthen your new relationships Here are some ways to go about strengthening a new relationship...events for one organization or go to the same conferences/meetings over and over to get to know the regulars...often the easiest way to build relationships and strengthen them over time.  Things to avoid when following...starfish shape. You can ask me about the different arms of this starfish sometime, but as of yesterday... was only one other person in my class who had children. When I had my baby, we became closer. She supported...really understood what I was going through. (Parenthetically, when I called the lab to ask my benchmate ...come across your contacts over the years in many different ways. This leads me to my last tip: It’s usually...

...plasmid cloning, and viruses. References Baltrus, David A., and Karen Guillemin. "Multiple phases of ...unique to the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...Various factors promote natural transformation in different bacteria such as growth phase of the cells (Baltrus...plasmid, the genes on the plasmid can give them different phenotypes. For example, this plasmid encodes ...cell. Once inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages ...a common tool used by scientists to introduce different DNA sequences of interest into a bacterial cell...through cell division. You can find all kinds of different lentiviral and AAV plasmids as well as ready-to-use...

... activate different G proteins, work in different neurons and neuroglia, and have different effects on...internalization of two different receptors- angiotensin II type 1 receptor and the β2 adrenoceptor. GPCRs in chemogenetics...Addgene’s chemogenetics guide.     References and resources References Kroeze WK, Sassano MF, Huang X-P,...These receptors can be activated by an array of different ligands, including hormones, neurotransmitters...is estimated that there are approximately 950 different human genes encoding GPCRs (Takeda et al., 2002...

...St Pierre is currently a graduate student at Northeastern School of Nursing and currently works as a registered...low-income communities to educate parents about resources that can help their children thrive academically. Rose...helps you find internships and scholarships in different subject areas!    2. Resume and cover letter...your relationships to the person as well as their current sphere of influence. In other words, the longer...the better, and their leadership status in their current field of expertise augments their authoritative... while you make the ultimate case: why you? 7. Strength in numbers Encourage your close peers to come ...C. St Pierre! Roodolph (Roo) P. St Pierre is currently a doctoral candidate in the Chemical Biology Program...

...international mentoring conference held at the University of Valencia (Spain). At this conference, mentors and mentees...international mentors. Universities from five different geographical locations in Spain were enrolled:...mentees held scheduled monthly progress video conferences and had real-time correspondence via e-mail throughout...Mentorship Program and the International Mentorship Conference In addition to a pre-program self-assessment ... exchanged impressions of the IMP experience. Conference highlights included remarks by the Spanish Secretary...community overseas. Official representatives from different Spanish universities and high level public and...their support and hope for the IMP program. The conference also honored IMP Mentors & Mentees with awards...

... words, they were leaving these meetings with different interpretations of what we discussed and decided...scientists learn how to communicate their work differently based on their audience. How you talk about your...at a poster is going to vary depending on the conference you’re attending and who approaches you. You ...with professionals from other backgrounds at conferences or networking events. And if you’re a wet lab...fellow team member. But, at Addgene, we use seven different types of “plates” (Fig. 1). When you have to tell...certainly wasn’t the only problematic word. How many different things do you simply refer to as a “tube”? We... they could in fact be interpreted in several different ways depending on the team. When Tony, our corporate...

...everyone at this specific conference, and I was able to include and exclude different modules for this workshop...something about it. Building a diverse team with different talents and expertise Fortunately, the eLife ambassadors...realized that holding the next one at a scientific conference could potentially enable reproducibility practices...it physically to instruct the workshop at the conference, but because each contributor made the materials...open and modular nature of the workshop content: different parts could be mixed and matched to serve a specific...working with communities, not for them - Another strength of this collaboration is that it is driven by ...the lookout for upcoming workshops at future conferences this year. Workshops are confirmed for the Evolution...

...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three distinct colors...orthologs is key: since each ortholog requires a different PAM sequence, a gRNA designed for one dCas9 should...specific for telomeric sequences and showed that different fluorescently labeled dCas9s are efficiently directed...target sequence. They succeeded in labeling two different pairs of chromosomes using gRNAs specific to sequences...specified by PAM sequence, each color requires a different Cas9 ortholog, and the target sequence must be... green, and red. In contrast, a gRNA with two different hairpins will produce a secondary color cyan, ...sequence (as noted in the table). gRNAs with two different hairpin sequences produce yellow, cyan, or magenta...

...24, 2025 by Alyssa Shepard.   References and Resources References Beumer, K. J., Trautman, J. K....Cong worked with Addgene Senior Scientist, Matt Ferenc, to organize the CRISPR forum's most popular questions...double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will... like the wildtype Cas9 does. There are a few references for the double nickase system, including one ...system probably have better specificity. A good reference is Ran et al. (2013). As for target selection...most guides would work, but they sometimes have different efficiency). And it is best if the cut site is...for one well in 24-well plate. A good example/reference is Ran et al. (2013). In the case where you ...

...collaborate with specialists in different fields that can provide different insight into an area - not just...you are looking to move to a different university, perhaps in a different country, choose where you go...collaborators It’s a good idea to move around to different labs during your career; you learn new techniques...I hope to discover. This section may make the difference in whether you get funding or not; applications... grants too There are actually quite a lot of different pots of money available for researchers, so look...gained in writing small travel grants to attend conferences and a seed funding grant for a proof-of-principal...mechanisms of cell division. Recently Seán got the entrepreneurial bug and has set up an ELISA kit company on ...

...concentrating a viral stock. Different titering methods can reveal differences in physical versus functional...negatively-charged membranes (Davis et al., 2002). Polybrene is one such cationic reagent that is commonly ...efficiency by 10-fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and ...be stored in single-use aliquots. Never use a polybrene solution that has been thawed more than once. ...in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and has...packaging cells for infection If your transductions aren’t working, you can check if virus is being generated...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...

...) signal. While many studies focus on promoter strength as a determinant of gene expression levels, the...these will not be detailed here, but additional references are provided at the end.  Nearly all common bacterial..., and III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ...that each eukaryotic RNA polymerase terminates differently. Polymerase III, for example, relies on a specific...Rho-independent termination found in prokaryotes. This is different than Polymerases I and II, which both require ...factor dependent, Polymerases I and II employ different mechanisms to terminate transcription. Polymerase...and/or use weaker poly(A) signals like BGH.  References Peters JM, Vangeloff AD, Landick R. Bacterial...

... Evaluating Gene function by RNA interference (RNAi) RNA interference (RNAi) is a powerful tool for performing...genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to knockdown expression of specific gene...morgani, and Phagocata gracillis. There are 11 different genes cloned into plasmids to investigate for ...WISH technique. The expression patterns of 11 different planarian genes (all of which are available through...phenotypes of RNAi knockdown treatment should be apparent after three dsRNA feedings. Some of the phenotypes...and if you make new plasmids for investigating different planarian genes, we encourage you to deposit them...researchers can easily investigate in planarians. References 1. Alice Accorsi, Monique M. Williams, Eric J...

... to run an ELISA in your lab!   References and resources References Boguszewska, K., Szewczuk, M., Urbaniak...Antibodies are used in many different experiments that require scientists to detect proteins in their...enzymatic reaction. Types of ELISA There are four different types of ELISAs, which differ in the way the antigen...antibody and the detection antibody recognize different epitopes on the target protein, so they do not... for which multiple antibodies that recognize different epitopes are available; this type of ELISA is ... are coated with another protein — known as a reference antigen — that can also bind to that antibody....antigen as the other types of ELISAs require two different antibodies. As mentioned above, it can also be...

...you can quickly test that part in different contexts and plug different characterized parts into a TU! What...works on genetic code expansion.  References and Resources References Weber E, Engler C, Gruetzner R, Werner... then chances are you have been in the cloning trenches yourself. Modular cloning, or MoClo, is a systematic...be assembled into many combinations. Even with current technologies, it can be extremely hard to predict...nature of MoClo allows users to test a variety of different orientations and components within the system ... a great tool to deliver many combinations of different candidate genes at once to identify the best cocktail...and gene editing that use MoClo, for which many different MoClo kits have already been established. Keep...

... transposition.  Target site: Different transposons insert at different DNA sequences or target sites....specific locations in the genome.   References and Resources References Feschotte C, Pritham EJ (2007) DNA... inserted into new target sites. Created in BioRender.com.   Class II TEs: DNA transposons Class ...a target site duplication (TSD). Created in BioRender.com.   Autonomous vs. non-autonomous TEs You...commonly used in the lab While there are many different types of transposons, DNA transposons are most... vectors.   Let’s take a closer look at three different transposon systems adapted for use as research...(Sandoval-Villegas et al., 2021). Target site preference and host range are important to consider when...

...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria.... Mobile-CRISPRi from the labs of Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined...plasmids for transfer to recipient cells during a tri-parental mating scheme. Mobile-CRISPRi integrates downstream...following IPTG induction of conjugation genes and bi-parental mating. Following excision from the B. subtilis...a recipient bacteria strain of interest by tri-parental mating. Donor bacteria contain a chromosomal copy...components is transferred to recipient bacteria by bi-parental mating. The ICE integrates into the trnS-leu2 ... of guides, promoters, selection markers, and different versions of dCas9. Following integration, Mobile-CRISPRi...

...overlapping markers. Compensation/reference control(s) Compensation or “reference” controls are samples stained...spectral flow cytometry are more alike than they are different, there are a few aspects in which spectral flow...complex mathematical algorithm and single stained reference controls to distinguish one fluorophore from another...   Figure 3: Similarity index across 12 different colors on a three laser spectral flow cytometer...conventional or spectral flow. In spectral flow, reference controls tell the software to look for that exact...is not the same as your single color control. Reference controls: Beads or cells? Compensation beads are...new panel on a spectral cytometer, you prepare reference controls of both beads and cells for a single ...

...leverages the strengths of PRIDICT/PRIDICT2.0 while accounting for some of the inherent inaccuracies of...focus on gene editing techniques. References and resources References Akhtar, W., De Jong, J., Pindyurin...means we could design hundreds to thousands of different pegRNAs for each desired edit, it simply becomes...many more. Once we had our datasets with all the different features and editing efficiencies, we could train...offline via GitHub. The model provides users with different designs of pegRNAs, ranked by their PRIDICT/PRIDICT2.0... predictive power is sometimes limited across different targets due to variations in chromatin context... Our recent studies (Mathis et al., 2024) and concurrent research by the Shendure lab (Li et al., 2024...

...subtypes of A–G, which come from different species and will have different compositions of Cascade. Although...through G, with different compositions of the Cascade complex. Created with BioRender.com.   The Cascade...degradative power in the future! References and Resources References Loeff, L., Brouns, S. J., & Joo, ... archaea. Class 1 is further broken down into different types, with Cas3 systems making up type I, the... complex from CRISPR type I-E. Created with BioRender.com. Thanks to this complex, developing Cas3...of the Cascade-Cas3 mechanism. Created with BioRender.com.   The reeling mechanism The reeling mechanism...

...discomfort of the mice, and restraint method. Many different substances can be injected, including cells, drugs...injection routes in mice. Image created with biorender.com.   Needle techniques Intravenous Intravenous.... They are a popular choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal...sciatic nerve. Mice likely experience some muscle soreness after injection, like you might after getting ...as both procedures target the lungs. The main differences are that intranasal is easier, less invasive,...be flexible or metal, depending on researcher preferences. It sounds complicated but has a pretty low learning... to 10 mL/kg Intermediate Resources and references Additional resources on the Addgene blog Mouse...

...yours does and then use that time! Different platforms have different quirks, so remember to practice on...that requires its students to do rotations in different labs before selecting an advisor, it is always...points and keeping them at-hand, you can quickly reference your questions about the PI’s research or the ...the name of the award you won for your conference presentation. It can help soothe your nerves and make ...promoted during COVID-19, online meetings with current students, faculty, and staff have become a new ...of just the program requirements: by talking to current students about the work environment and social ...using the virtual world to your advantage can strengthen your application and help you decide if a program...

...Organizing a science conference for the public How does one organize a public scientific conference? As a member...first-time event called DayCon. DayCon is a one-day conference aimed at the general public that provides an ... commitment of SITN members. SITN and DayCon Currently in its 16th year, Science in the News has two ...scientists and the public and help make science more transparent. Second, SITN helps graduate students develop...graduate student at Harvard, who wanted to provide a conference-style experience that was accessible to the general...picked a theme (Bio-Medley) to showcase SITN’s strength in the biomedical sciences and decided to offer... make DayCon a yearly event and to expand the conference to accommodate a larger number of attendees. ...

... of antibiotics, but the field is currently undergoing a renaissance. Yosef et al. designed a two-phage...lysogenic phage, rendering the population sensitive to antibiotics. Created with BioRender.com. Phages ...swift, as few as five years after introduction. Current antibiotic strategies are nonspecific - they harm...defense against bacteria. Created with BioRender.com.   The problem of antibiotic resistance ...alter an individual’s native, beneficial flora. Currently, there is no antibiotic strategy available to ...mixed population, represented here by cells of different colors, specifically targets bacteria with the...ground in the fight against bacterial superbugs. References 1. Citorik, Robert J., Mimee, Mark, and Timothy...

...the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer have been extensively...compromised either due to age, infections, or other strenuous biological changes. In contrast to the acute immune... cytokines like TGFβ and IL-6. However, when strengthened through efforts to boost the activity of the... most commonly used immunotherapeutic agents. Currently the FDA has 12 approved monoclonal antibodies .... Sipuleucel-T by the Dendreon Corporation is currently the only FDA-approved DC therapy for prostate ...only just touched the surface of immunotherapies currently under study. Check out this review for an in-depth...manifestation to their specific interactions with different cell types. A thorough understanding of the genotypic...

...soils. Dendrobatidae (Lauren O’Connell)  Poison frogs are great for studying parenting strategies and make...land. Current tools are all for Xenopus (aquatic frogs), but there are few genetic tools currently for ...Genomic Tools) Program and I met scientists from 21 different labs who clearly also share this wonder of the...convergent evolution because electric organs arose in different branches of the evolutionary tree at least 6-7...editing in the brain allows the squid to create a different protein. Did you know that most cephalopods die...behavior. For example, two similar species have very different social behavior. One is monogamous and one is ...new tools being developed and shared for many different organisms. Do you have a favorite organism you...

...educational training, and conferences, and synthesizing this knowledge into a coherent narrative that gives...best with diverse teams, to leverage the preferences of different personality types, and to devise strategies...biotech clients. In this realm, there are a few different kinds of challenges that consultants get called...companies. Medical Affairs groups work on building awareness among physicians and scientists about clinical...consulting firms coach and counsel employees on the different personality and work styles (e.g., Myers-Briggs...prepare you for a successful career in several different fields. While some people continue to enjoy the...BioCentury can be an engaging way to learn about the trends and workings of the healthcare industry. Finally...

...doping to forensics, DNA sequencing to drug testing. You can find Nick on LinkedIn. References 1. Skloot...Expression Attenuates Proliferation and Invasion in Renal Cancer Cells, Marker-independent identification ...the cell lines deposited with them and used by different researchers are free of contamination and match...from this bank. I am often asked, “What is the difference between materials available from commercial repositories...the same cell line from a colleague?”. The main difference is the QC and validation carried out by the commercial... The same test is so good that it is used as a forensic technique (aka DNA fingerprinting) that can place... produce a new batch once you’ve passaged your current batch too many times or you suspect there is contamination...

...going to cite hundreds of references) and it’s better to keep your references organized from the beginning...the thought of writing a review, or if you’re currently stuck trying to write one, hopefully this post...criteria to shape the document. Get and use a reference management program (e.g., EndNote, Papers, Mendeley...Mendeley, etc.) You’re going to be managing a lot of references. I cite as I write, meaning I use the software...in real time as I wrote, so each statement was referenced even in the roughest version of the draft and...that people can go back and read the original reference for the data. For example, we summarized how frequently...place to get a sense of the similarities and differences among proteins in the class. Offer your perspective...

...for other antibody topics.     References and resources References:  Björck L, Kronvall G (1984) Purification...elute and measure the target. However, the two different approaches allow for optimized strategies for ...amounts of target proteins or antibody bonding strengths.    . Steps of an IP: 1) Protein extracts... appropriate for most situations.  A slightly different method is desirable if the antibody is expected...Protein G have an affinity for immunoglobulins, the strength of the interaction varies between antibody host...than those of agarose beads, despite the size difference. Magnetic beads are particularly advantageous...many factors, including salt concentration, ionic strength and pH, affect binding the lysis buffer may need...

... can separate the DNA strands. Created with BioRender.com. Once DnaA breaks the local hydrogen bonds...Prokaryotic chromosomal replication. Adapted from BioRender template. Since DnaA binding is necessary ...proteins also contribute to this regulation in different ways: methylating newly synthesized DNA, blocking...numbers can allow RepA to link iterons from two different plasmids, forming a pair of “handcuffs” that blocks...RepA proteins form complexes between iterons on different plasmids in a process called "handcuffing," which...folding, blocking replication. Created with BioRender.com. Plasmids under stringent control of replication...was updated by Emily P. Bentley in June 2024.  References and resources Further Reading Quiñones-Valles...

...host species. Created with BioRender.com. How are shuttle vectors different from other plasmids? If ...protects against any of them. But different aminoglycosides work on different host species: kanamycin targets... unrelated host species, even from completely different kingdoms of life. That’s convenient for researchers... Shuttle vectors have to address several big differences in the ways vectors are used across species: ...resistance gene on your vector might require two different selection molecules! It all depends on how the...Auxotrophic selection Another option is to use a different method of selection entirely. Auxotrophic selection...host range,” these terms are typically used in different (though overlapping) ways. Broad host range plasmids...

...virus:  Lowering cytotoxicity. Different envelope proteins have different levels of cytotoxicity and the...Altering sensitivity to serum. Different pseudotyped viruses are differentially inactivated by nonspecific...Table 1 (Cronin et al., 2005)     References and Resources References: Crawford KHD, Eguia R, Dingens AS...protocol for generating lentivirus, but you use a different envelope glycoprotein instead of the wild-type...review articles to find tables and information on different viral membranes, tropism, and advantages and disadvantages....084 Li Q, Liu Q, Huang W, Li X, Wang Y (2017) Current status on the development of pseudoviruses for ...

...expect, different companies produce different products and therefore need people with different areas of...: vast practical lab expertise, experience in different scientific fields, and knowledge of how to troubleshoot... as: “Can I use this product under conditions different from those specified on its data sheet?” or “Do... questions customers ask, and there are also differences in customers’ personalities and their expectations...do jobs in technical support have? There are different names for technical support roles. Examples are...distinction between these job titles, they may be different regarding the levels of lab experience and scientific...communication skills by working with people from many different backgrounds and realizing that the same approach...

...are communicating across disciplines or doing different tasks. But these challenges can be exaggerated...equal opportunity to contribute. Using video conferencing tools like Zoom or Google Hangouts is a great...great way to include researchers from different locations. Having virtual face-to-face meetings as opposed... collaboration includes scientists working in different time zones, it’s important that everyone can access...project at the International Cancer Immunotherapy Conference in Frankfurt, Germany When funding and circumstances...techniques are being performed the same way at different sites. I did a lab exchange as part of the collaboration...allowed me to investigate my protein of interest in a French clinical cohort, neither of which I would have ...

.... The plasmids within it contain a variety of different promoters followed by leader sequences (encoding... at the BIOFAB showed that expression of many different genes was predictable based solely on the promoters...BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP bicistronic reporters ...or protease sites to remove the epitope tags. Different plasmids in the kit can be used to purify proteins...prokaryotic expression in recent years, with many different systems now available. However, it can be difficult...ORF clones to allow testing and comparison of different cell-free expression systems; the Cell-free expression...bind specific RNA bases. By swapping out these different modular repeat segments, you can create a PUF ...

...what we need to know in order to even assess the current state of research became more clear throughout ...that, even though postdoc associations across different universities are trying to compile more comprehensive...path,” but individual institutions can have many different names for the positions commonly thought of as...the Starting Salary for Postdocs to $50,000 (the current median postdoc starting salary as reported by Paula...Student Career Outcomes University websites do not currently publish career outcomes for PhDs or postdocs. ...achieved. Of note, it was highlighted during the conference that just increasing funding for academic research... At Addgene we try to do our part in boosting awareness of various career opportunities through our career...

...microscopy and beautiful imaging. References and Resources References  He, Y., Zhang, T., Sun, H. et al...killing the untransformed seedlings or callus (undifferentiated plant cell culture), which is great for a ...desired plants! PCR or fluorescent markers are currently used in these situations to select against CRISPR...such as cacti and beets. Betanin belongs to a different chemical family than the red/purple plant pigments...it for selecting transformation events and am currently planning to use it to simplify my work with an...(Bottom panels) RUBY expression can vary from different transformation events likely due to insertional...lower expressing RUBY plant is a pale pink in transparent tissues, or in green tissues is slightly darker...

...Why the HEK not? References and Resources References Shaw G., et al., Preferential transformation of ...determined it is most likely adrenal in origin. (Anatomy refresher: the adrenal gland is next to the kidney...were isolated from is still a mystery, but it’s currently assumed that the cells came from an aborted fetus...genetically female. HEKs are generally cultured as an adherent monolayer, though derivatives have been adapted...have some neuronal qualities.) While HEKs probably aren’t a great model of kidney cell biology, there are...

...of many complex cellular behaviors, such as differentiation or metabolism. Our attempts to manipulate these...genes or to introduce heterologous genes, but the trend of research has shifted in recent years toward tinkering...system for multidirectional control The CRISPR interference (CRISPRi) model consists of a catalytically ... multiplex control via scRNA design is readily apparent in metabolic engineering. The authors elegantly.... cerevisiae by using three RNA programs to differentially alter the levels of five genes that encode ...sequentially rerouted the metabolic flux to generate differently pigmented derivatives of violacein. Using dCas9...that will still recognize guide RNA but with a different dCas9 binding module, alternative sets of transcriptional...

...available jobs, fields, and trends “Never Leave an Empty Seat” at conferences and seminars, as this is ... path to professorship. But recently a downward trend in funding and space in academic research labs has...traditional academic setting and to shape and strengthen interests and skills. Although the word “extracurricular...potential EAs and which professional skill sets are strengthened. This short list highlights the ubiquity of ...particular field of study Assess your interest in a different field Outreach activities (e.g. Volunteer...Marketing Communication Business models Conference or club organizer Management Leadership...know where graduate students and postdocs are currently getting hired? Let us know what topic you'd like...

...seeing adoption of the technology in different labs for different applications.” Li’s team noted that ... genetic diseases, and several treatments are currently being tested in clinical trials. It is speeding...cells of patients with cystic fibrosis (8).   The difference with CRISPR/Cas9 in comparison to other genome...for selection and optimization. In addition, the current work in developing anti-CRISPR proteins aid the...possible through the use of nanoparticles. [6].    References 1. Zhang B. 2021. “CRISPR/Cas gene therapy.” ...at genome editing with CRISPR-Cas9 technology.” Current genetics. https://doi.org/10.1007/s00294-019-01040...its Application in Genetic Diseases: A Review.” Current pharmaceutical biotechnology. https://doi.org/10.2174...

...of the program beneath it. You can jump to the different time stamps in the description to watch any sections...precisely. You can find a great discussion of different types of replication in Nosek and Errington 2017...another’s reagents with the confidence that they aren’t simply doing an experiment wrong. However, even... a plate or running a gel have intricacies that aren’t always mentioned in static written protocols. It...scientific findings. This makes sense on many different levels - it’s hard to process large amounts of...to get a publication (for better or worse, the currency for success in academic research). Initial positive...review our experimental designs and we need to be transparent about the reasons we alter these designs if we...

.... Unfortunately, there really aren’t any effective off-target reference controls. Off-targets and your...they do arise. Happy CRISPR’ing! References and Resources   References Rabinowitz R, Offen D. Single-Base...effects should impact your experimental design differently. Yes, it is always better to reduce off-target...mutation, off-targets must be quantified slightly differently. For a comprehensive approach, RNA-seq would ...acknowledged as a potential limitation in a study. Strength in numbers If you assess your off-target frequency...to control for off targets as well as clonal differences. If you are performing CRISPR library screens... as a transfection or transduction control, a different gRNA would have distinct off-target sites, eliminating...

...isolated from different organisms and engineered Cas9 variants that were able to target different PAM sites...activity of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., 2018)...found in other species can recognize a variety of different PAM sequences. Researchers have also used experimental...specificity can target more sites. Created with BioRender.com. Specificity Specificity is the assessment...relative to eSpCas9 and SpCas9-HF across a number of different sites. Using a combined random and directed mutagenesis...choosing a Cas protein for your experiment. References Hu, Johnny H., et al. "Evolved Cas9 variants ...

...performance under different imaging conditions (e.g. different emission filters) or different set-ups. Many...uptake by cells. To correct for this variation, a reference fluorescent protein that is expressed at an equal...be used. An assay developed in our lab uses a reference protein that is translated from the same open-...between the fluorescent protein of interest and the reference protein (Goedhart et al, 2011) allows for a precise... a set of fluorescent proteins (with the same reference protein) under identical conditions, a ranking...for co-expressing mTurquoise2 with mVenus as a reference protein is available from Addgene. Another way...

...Nano-lantern technology. Nano-lanterns contain a Renilla luciferase variant fused to an FP; when supplied...Solving the problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently... first Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent...resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite the fused...technique’s versatility and adaptability to many different contexts. Improving Nano-lanterns In 2015, Takai...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially... for excitation, you instead have to rely on differences in emission wavelength. Filtering the emission...

...imaging for advanced monitoring of different factors, in different or the same cellular location, all ...covered with packaging plasmids! References and Resources References  Bhalla N., Jolly P., et al. Introduction...biosensors provide quantifiable readouts for levels of different chemicals, hormones, proteins, etc., that you ...and zinc. These ion sensors can be targeted to different cellular locations or tissues depending on the...acceptor of a FRET pair on actinin will produce different signals based on changes to the structure of actinin...organisms, but AAVs make it possible. In AAV systems, different serotypes can be used to affect or determine viral...

...skills that were quite different from mine. Learning to bridge those differences was a pivotal moment in...technical problems in the lab, management involves a different skill set: mentoring, setting strategic goals,... day-to-day details. Ignoring individual differencesManaging a team isn’t “one size fits all”. Each team...team member has unique motivations, strengths, and learning styles. For example, during my time as a lab...autonomously. Failing to recognize and adapt to these differences could have led to frustration, micromanagement...cohesive, productive environment. For example, in my current role managing a cross-functional team on a biomarker...team meeting to highlight their complementary strengths. By encouraging alignment on goals and clarifying...