...CRISPR technology has brought drug and genetic screens together and expedited the drug discovery pipeline...Alternatively, a disease model can be generated, and drugs screened for a change in pathology. If the disease pathology...accelerate the drug pipeline. High-throughput CRISPR screens can be used to inhibit, activate, or knockout many...or an entire pathway of possibilities greatly increases the odds of finding a quality drug downstream....identified several candidate inhibitors of gene X in a screen. You generate a gene X knockout in a cancerous ...

...isolated and screened. Depending on the marker, traps can be used as high-throughput screens to introduce...models. Why traps are useful Traditional genetic screens employ various on/off switches to measure the function...circumstances, trap methods may be the preferred screening option. Promoter traps Promoter trap systems ...101: Using Transposons in the Lab Plasmids 101: Screens vs. Selections Plasmids 101: Knockout /Knock-In...

...activities (1). She has developed a fast endophyte screening system and plans to employ the light‐inducible...expert. In order to preserve Cassava yields and to increase the potential for this drought tolerant plant,...significant proteins. In this project she aims to create transgenic yeast as an alternative source of essential...plasmids. The recombinant Cry proteins will then be screened for toxicity against local insect pests. By developing... use of chemical pesticides may be avoided or decreased. Biosensors for herbicide detection Dr. Rodrigo...

...CRISPR screens symposia outlined the fantastic work this Broad Institute lab is doing creating well-optimized...well-optimized CRISPR screening technology. Addgene got several shout-outs for making popular screening libraries ...presence in the microbiology community. For our first concrete steps into this field, we’ve curated microbiology...

...pENN.AAV.hSyn.HI.eGFP-Cre.WPRE.SV40 AAV2 Recombinase James M. Wilson New serotype pENN.AAV.hSyn.Cre.WPRE.hGH AAV2 Recombinase....GFP-Cre.WPRE.SV40 AAVrg Recombinase James M. Wilson New serotype pENN.AAV.CamKII 0.4.Cre.SV40 AAVrg... viral prep AiP14825: pAAV-AiE0441h_3xC2-minBG-iCre(R297T)-BGHpA (Alias: CN4825) AAV-PHPeB Recombinase...

...cobalamin fusion to the riboswitch results in increased fluorescent signal. Prior to the development of...developing a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer...the red end of the spectrum (above 600 nm) and increasing the brightness. Arne Fabritius and colleagues...Listen to the base editing podcast segment The increasing popularity of single base-editors has resulted...cytidine deaminases like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC...

...large-scale functional screen in bacteria. Sensor variants were then refined by a secondary screen in rat hippocampal...protein (CREB) in living cells. “Now we are making this really good,” Griesbeck said. “We know CREB is really...10.1038/nmeth.2773. (PubMed) Friedrich et al. Imaging CREB activation in living cells. J Biol Chem. 2010 Jul...

...photoconverts predominantly to the anionic form, causing increased fluorescence when excited with 488nm blue light...which allows PA-GFP to exhibit up to a 100-fold increase in fluorescence when illuminated with blue light...performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry (E26V / ...photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't become...the absence of photoactivation resulting in an increased signal to noise ratio. PA-FPs with higher contrast...super resolution microscopy, PA-FPs have helped create several new imaging methods, including photoactivation-localization...

...protein secretion, making harvesting recombinant proteins a cinch. This is because it doesn’t secrete its ...of producing high protein animal feed. Due to increasing prices of methanol and advancements in other ...proteins, making the recombinant protein its major secretion. Beyond sheer protein production, K. phaffii is...Organisms Plasmids 101: Yeast Vectors Tips for Screening with the Yeast Two Hybrid System Additional...

...purple) and Gram negative (red/pink) staining. Image credit: Mark Perkins https://www.flickr.com/photos/occbio...charged iodine is added, it interacts with the CV+, creating a complex within the peptidoglycan layer, staining...peptidoglycan layer, trapping the large dye complex created by the interaction of CV+ and I- in the thick multilayered...positive and negative classification system is incredibly helpful in visualizing differentiations between...What is a plasmid? Plasmids 101: Blue-white screeningPlasmids 101: Common lab E. Coli strains  ...

... lack of credit for small contributions. We strive to eliminate these two barriers, creating a community...with others than by oneself that prompted us to create Crowdfight.   Figure 1: Crowdfight is a ...collaboration between researcher and expert; 4) Credit all contributions.   Our volunteers are extremely...collaborations can be easily started and easily credited. And nothing stops these one-time favors from ...

...roughly 200 cells. Increase the titer to 1013 resulted in a roughly 6-fold increase in expression [3, ...Figure 2]. Note how expression increases exponentially with an increase in titer. That means that once...to get more AAV in contact with the cell is to increase the AAV concentration. In this flowchart, you ...

...primers to create 200-300 bp sequencing products using PCR. A diagram of the product we created for the ...plasmid. However, lots of library DNA is needed for screening experiments, so the scale of the DNA prep is larger...rank the plasmids by abundance, scaled to 1, to create a “cumulative wealth distribution” curve, or Lorenz...wishes you luck in your future library use and screening, and if you have any tips, input, or stories of...

...addition to increased brightness, they also found that the signal to noise ratio can be increased from 2-fold...previous CRISPR-based loci visualization tools by increasing gRNA stability. First, they began with the Broccoli...and went through subsequent design iterations to increase stability and minimize misfolding and recombination...label two loci simultaneously, Ma and Pederson created another sgRNA containing the PP7 hairpin sequence...

...resulting in a decrease in luciferase activity. This experimental protocol can readily screen miRNAs for ...Peak emission (nm) Size (kDa) Substrate Cofactors Secreted Luminous shrimp(Oplophorus gracilirostris) 455...cell membrane and deliver the substrate; however, secreted luciferase or the use of alternative substrate...pathways can be investigated in a high-throughput screen using luciferase activity as a measure of signaling...

...that approach seems to create competition for your own spot. True! But you create a network of friends,... and simplify access to academic resources, we created STEAMid, for a nationwide database of  STEAM internships...with your question(s). This precautionary step increases the likelihood of acceptance to the internship... following graduation. While these suggestions increase your competitive edge, they alone, are no guarantee...

...the potential for increased speed and efficiency through which knowledge is created and shared. Can you...platform. Expanding further beyond the US and increasing Addgene’s international presence would potentially...information, making it available to everyone, that creates more breakthrough and innovation. With any kind...science regulatory law. My role at Suffolk is to help create new programmatic content as well as help our students...

...sequence desired. This technology allows you to create a pair of designer proteins that will introduce... transfection or electroporation. All you do to create a protein that binds those 15 base pairs is take...genome editing tools allow you to do is greatly increase the efficiency of the genome editing process. ...for investigators without a lot of expertise to create zinc finger nucleases that will bind desired sequences...in terms of the target site. Also, any time you create double-strand breaks, there is a possibility of...

...Institute has created hundreds of thousands of plasmids, using both mass production genome wide screens and the...vector. The time it takes to make a plasmid could increase dramatically if you are piecing together several...at Addgene, the consumables cost per stab has increased 50% since 2019, while overall inflation of USD...equal to the $155 spent on reagents. Five ways to decrease plasmid production time and cost If you’re short...

...visibility is increasing internationally, thanks to evolving outreach tactics, increasing business development...they see the value in depositing and they are increasingly seeking us out on their own to share reagents... We were and remain ahead of most companies in creating a welcoming, inclusive workplace for everyone....about their dedication, excellent intentions, or incredible drive to succeed.  Thanks to all the Addgenies...