... the field of grants. What is a grant professional?  In the past 5-10 years, the Grants Profession has...collaborations, and commitments to and proof of compliance with the grantor’s requirements as well as state... for grants, the amount of grant funds available and the increasing regulatory process for grants has ...grant consultants who offer grant training, mentoring and coaching services, including Smartegrants, to ...choice exam.  The bottom line on being a grants professional The Grants Profession is in a rapid growth phase... Inc., a national grant consulting firm, and Smartegrants, a grant professional development firm The path...established grant professionals is learning to succeed in grants through trial and error, through winning and losing...

.... Now is the time to make bold statements. While some of the statements may be slightly far reaching, ...consider the reviewer's comments. Whether the proposal is funded or not, the comments help gauge room for ...government and academic institutions. New graduate students may assume that their advisor alone applies for... allot some if not all grant duties to trusted students. For example, if the proposal call aligns with...write a grant!”, “I don’t have time for this if he wants data by next week!”, and “This is his job!” filled...various skill building opportunities that graduate students and postdocs should actively pursue. Proposal ...italic, or underline font formats helps these statements stand out from the rest of the text. Specific...

...single 96-well plate for HDR that satisfy all requirements. To alleviate this bottleneck, we began building...whole or in part. For example, you can analyze experiments designed outside CrispyCrunch, or you can input...step of mutating guide sequences in a way that prevents re-cutting but does not disrupt gene expression...that worked with NGS data were: Crispresso CrispRVariants AmpliCan We chose to adapt Crispresso by Luca...re-create a match for the active guide. With these constraints in mind, we implemented the following algorithm...Design donor RNA Design primers Review and order reagents Perform your wet lab experiment Analyze the data...high-throughput design and analysis of CRISPR experiments. It has certainly boosted our productivity in...

...(i.e. the plasmid complements the deletion). If the phenotype is corrected, students can be relatively... multiple enzymatic steps (5). In their experiments, students test whether or not proteins from the fission...extensive rearrangements in the S. cerevisiae lineage (7). In our class research project, students transform...in other experiments (Figure 2).   Complementation reveals conservation of function Students have used...Boston College. Students in the course work in teams of three in sections of 15 students. Within each section...also used to teach students about restriction mapping and western blotting. Students who participate in...authentic research experiences to undergraduate students (1, 2), but educators face significant challenges...

...levels and affect behavior. A fact to consider: Rodents can hear ultrasonic frequencies (> 20 kHz, produced...

...around the globe, we’re now sharing over 100,000 reagents including plasmids, ready-to-use viral preps, ...

.... Both awardees emphasized the contribution of students in their labs and the importance of the strong... what can be done with this technology to help patients. Dr. Stoffels and others in the audience discussed...technologies. Addgene is proud to be helping get CRISPR reagents out to academic labs. It is our hope that robust...

... cloning efficiency measurements. We performed additional efficiency measurements using similarly sized...number of transformants); the greater the number of inserts, the lower the number of transformants. To compensate... Table 1). These improvements (5- to 12-fold increases in the number of transformants while maintaining...reaction volumes were 20 μl (1, 12 fragments) or 25 μl (24 fragments). The future of assembly DNA assembly...Assembly is commonly used to assemble 2–10 DNA fragments in a single “one-pot” reaction to form complex... site is no longer present, allowing multiple fragments to be assembled in an ordered, pre-determined ...background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion of the...

...CRISPR applications in plants.     Transgenic yeast as source of dietary supplements Dr. Mrinalini Nair,...well as developing yeast as source for dietary supplements! Development of a bacterial expression system..., bacteria, and fungi that reside within living plants and that produce compounds with various antimicrobial...More Optogenetic Tools Like pDawn  Making cassava plants resistant to cassava mosaic virus Dr. Fiona Robertson...make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy DNA viruses, similar to...interested in applying gene technology to bacteria and plants. Her research involves cloning cry genes from the...against local insect pests. By developing biological agents such as insecticidal proteins, the use of chemical...

...responses in patients. They were able to tweak this assay to study SARS-CoV-2 spike variants. “We created...circulating variants. This allowed us to create pseudoviruses that represent lineages and variants that are...we’ve seen the emergence of several SARS-CoV-2 variants coincide with increased vaccination efforts. So...effective are the current vaccines against the variants? Alejandro Balazs, a Group Leader at the Ragon... of current vaccines differed depending on the variants (Garcia-Beltran et al., 2021). Antibodies from...Moderna or Pfizer vaccine could neutralize many variants, including the B.1.1.7 (Alpha variant), B1.1.298...well as they could for the wild-type. However, variants such as the P.1 (Gamma variant) and the B.1.351...

...share these improvements? You could publish a paper that happens to include your improvements, or maybe ...the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem many...information, you’ll have the option to add additional documents. By uploading your protocols onto your plasmid... to your protocols when they begin their own experiments. Need to submit an updated protocol? Just email...them public for others to use, and add your own comments. Currently you can find protocols from NEB, OpenWetWare...storage and inventory, managing orders, tracking students and research projects, electronic lab notebooks...trials and pricing options for academic labs, departments, industry, and more to fit every lab's needs....

...Quantifying the abundance of thousands of protein variants simultaneously  With increasing sequencing power...individual’s genome. But when they find genetic variants, what does that mean? Previous research shows ...approximately 75% of all protein-coding pathogenic human variants are decreased in protein abundance, possibly due... due to misfolding. But measuring variants one at a time is time consuming and unscalable.  Melissa Chiasson...Washington sought to address the effects of genetic variants in a high throughput manner. She helped develop...a method that measures the effects of sequence variants on abundance simultaneously called VAMP-seq, for...protein variant library is monitored by fusing the variants to eGFP followed by an internal ribosomal entry...

... Taq buffer with MgCl 2 1 μl dNTP mix (10 mM each nt) 2.5 μL Forward Primer (10 μM stock) 2.5 μL Reverse...total + 10% (3.5μl) = 38.5μl dNTP mix (10 mM each nt): 1µl x 7 reactions = 7µl total + 10% (0.7µl) = 7.7µl...thousands of identical copies using a simple set of reagents and a basic heating and cooling (denaturing and...of the DNA segment to be amplified. When these reagents are combined in an appropriate buffer, a series... the newly synthesized strands. Materials List Reagents for each 50µL PCR: Thin-walled PCR tube Ice Bucket... on ice Set up a 50 μL reaction (Keep all your reagents on ice): 2 μL Template DNA (10 ng-500 ng) 5 μl...

...and their research. These researchers span four continents and research diverse topics including microbiology... University of Ibadan. She works with medicinal plants such as Launaea taraxacifolia (Willd), an annual...renewable sensor, a bacterium that can sense contaminants and emit light as a response. To create this...fluorescent protein transcription in response to river contaminants and transform these plasmids into a bacterial...geographical locations. In addition to providing reagents and educational resources for scientists globally... Check out our CRISPR resources and CRISPR for plants Bacterial Expression Systems Synthetic Biology ...

... also shorter than those needed for Cas9 (42 nt vs 100 nt.) However, the newly described non-CRISPR protein...Additionally, whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1 cleaves in a staggered...smaller than any of these options (2.7 kb with 24 nt guides), so it has the potential to eclipse any of...into AAV when you add in a gRNA and regulatory elements. To solve this problem, the Zhang lab has previously...Kleinstiver et al., 2016. Until recently, two major elements of Cpf1 characterization were missing: on-target...

...GRAB biosensors for neuropeptides VIP, NPY, CCK, NTS, UCN, SST, and CRF A tool kit of highly selective...Quantitative measurements of membrane charges A FRET sensor enables quantitative measurements of membrane...designed to specifically localize to cellular compartments or organelles. Addgene’s database includes many...that many of the articles linked below include variants with different binding affinities or negative-...imaging activity in neuronal populations and microcompartments. Nat Methods. 2019 Jul;16(7):649-657. Douglas...astrocytes (GCaMP6f), cytosolic Conditions and constraints for astrocyte calcium signaling in the hippocampal... Calcium Yellow protein calcium sensor jYCaMP1 variants (Constitutive or Cre-dependent) jYCaMP: an optimized...

...NTF4 neurotrophin 4 NT-4/5, NT4, NT5, NTF5 NTS neurotensin NMN-125, NN, NT, NT/N, NTS1 NUDT6 nudix (nucleoside...Antibodies produced by B cells activate immune components that also lead to the destruction of pathogens...

...mismatches in target RNA from bases 12-26 of a 30 nt target sequence. For the editing portion of the protein...sequencing, they confirmed A->I editing and found that 50 nt spacers increase editing efficiency but also increase...efficiency. They note that RESCUE is most active in a 30 nt guide when C or U base-flips are present across from...

...for a surprise! Whereas Cas9 generates blunt ends 3 nt upstream of the PAM site, Cpf1 cleaves in a staggered...doesn’t require a tracrRNA, crRNA guides are only ~42 nt long. Direct synthesis of these crRNAs should be ...be significantly cheaper than that of the ~100 nt crRNA/tracrRNA hybrid guides needed for Cas9 function...depletion assay to discover FnCpf1’s PAM sequence requirements. Cpf1’s preferred PAM is 5’-TTN, differing from...tracrRNA. Once they had determined the minimal elements of CRISPR-Cpf1, Zetsche et al. turned to characterizing...

...Sequence NGG CRAA (R = A or G) Spacer Length 20 nt 22 nt Temperature Range for Optimal Activity 35 - ... organisms: mammalian cells, yeast, drosophila, plants, worms, zebrafish, frogs, some bacteria; but not...s love of heat in two ways. The first set of experiments tested the ability of GeoCas9 RNPs to be incubated... following heat treatment. The second set of experiments tested the ability of GeoCas9 RNPs to cleave ... we haven’t thought of yet? Let us know in the comments below. And if you want to give GeoCas9 a try, ...