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...space to be oxidized and form disulfide bonds (Frenzel et. al., 2013). If you’re using a mammalian production...: The steps of rAb production. Created with biorender.com   Another method of harvesting uses magnetic...useful to compare recombinant antibodies to their parental hybridomas. Aggregation and storage Aggregation...our Antibodies 101 blog series.  Resources and references More resources on the Addgene blog Introduction...Purification Protocol Addgene's Antibody Guide References Trimmer JS. Recombinant Antibodies in Basic Neuroscience...: 10.1007/978-1-4939-8730-6_1. PMID: 30242676. Frenzel A, Hust M, Schirrmann T. Expression of recombinant...particle formation, characterization & rheology, Current Opinion in Colloid & Interface Science, Volume ...

...Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2 noradrenergic area of the caudal...helping everybody, helps everybody! References and resources References Dayton, R. D., Grames, M. S., & ...scientists under-informed with respect to how different serotypes, promoters, and injection methods are...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...particle aggregation and methods to prevent its occurrence during vector purification and formulation. Molecular...

... as fixing and permeabilizing.  References and resources References Brown-Harding H (2020) Fixation artifacts...previously hidden within the protein. Created with BioRender.   Permeabilization What is the point?  Antibodies...100, Tween-20, or saponin. Each detergent has different chemical properties that will impact the level...concentration.      Figure 2: Examples of different permeabilization strategies and when to use them...permeabilizing organellar membranes. Created with BioRender.   How do I choose?! Ok, so now you know some...https://doi.org/10.1177/33.8.3894499    Im K, Mareninov S, Diaz MFP, Yong WH (2018) An Introduction to...

...It was by serendipity that I got into the field of gene therapy, more specifically AAV-based retinal ...treatment, famously visited Congress to help increase awareness about the potential of gene therapy!). It would...is available. While only natural serotypes are currently used in clinical trials, a variety of engineered... AAV safety profile is excellent and further strengthens its position as the vector of choice for in vivo...and the development of personalized medicine. References and further reading 1. FDA approves gene therapy...the Light after 25 Years of Retinal Gene Therapy. Trends Mol Med. 2018, S1471-4914(18)30123-0. DOI: 10.1016... Karen Guerin...

...be identified by PCR later. The PNS presents a different problem: it’s spread throughout the body and is..., AAV.PHP.eB was used at a lower dose than it’s parent AAV.PHP.B: 1x1011 vs 1x1012 viral genomes (vg) ...comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids ...efficiency of AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for...   Cortex Striatum Cerebellum AAV.PHP.B (parental) 49 +/- 7% 36 +/- 8% 37 +/- 10% AAV.PHP.eB 69...Transduced   Dorsal Root Ganglion (DRG) AAV.9 (parental) 46 +/- 0.7% AAV.PHP.S 82 +/- 2% Table 2:... and supplemental figure 8 in Chan et al).  References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered...

...tools with a focus on plants.     References and Resources References Karmakar, S., Panda, D., Baig, ...TnpB protein, much like how PAM sites change for different Cas species. For instance, the TnpB from Deinococcus...Deinococcus geothermalis (Dge10TnpB) targets a different TAM (5′-TTAT-3′). The unique TAM requirements ...for plant genome editing using TnpB. The main difference between them lies in the choice of promoter for...tools for predicting efficiency of guides are currently unavailable, you can use TEEP — a tool based on...Engineering the Plant Genome Using CRISPR/Cas9 RNA Interference in Plant Biology: New Tools for an Old Favorite...

...regulated transcription units. Created with BioRender.com.   Planning makes perfect Our first tip,...vectors and parts you will be using during the different levels of your MoClo assembly. Know your process...dictated by four-base overhangs that are paired with different part types (e.g. 5’-GGAG and 3’-TACT flanking ...ensure that these would be interchangeable between different laboratories. Therefore, choosing acceptor vectors... Example Level 1 assembly. The choice between different Level 0 vectors depends on the four-base overhangs...C-terminal tag; TER, terminator. Created with BioRender.com. The Sainsbury Laboratory Golden Gate Cloning...you plate after each transformation and/or use different E. coli strains with higher transformation efficiency...

...scientists have since adapted it for a host of different proteins and small molecules from a variety of...needs. Herein, we’ll give you a rundown of the different types of ELISA, advantages and disadvantages of...in the sample. This figure was created with BioRender.com  Direct ELISAs A direct ELISA is the quickest...this method, the target antigen competes with a reference antigen for binding to a primary antibody.  While...typically users first coat the multiwell plate with a reference antigen and then they preincubate the antigen-...allow the unbound primary antibody to bind to the reference antigen. The more antigen in the sample, the more... means less antibody available to bind to the reference antigen, producing a weaker signal.  The major...

...colleagues reported that it is possible to reprogram differentiated cells into induced pluripotent stem cells (...benefit when recruiting patients, specifically children and patients with skin disorders for research ...OriP and induces amplification of the episomes concurrently with DNA amplification in the host cell. This...integration-free, karyotypically normal and are able to differentiate into various cell types of all three germ layers...Kushi) Mukherjee. Kusumika (Kushi) Mukherjee is currently a postdoctoral fellow at Massachusetts General...https://www.linkedin.com/in/kmukherjeephd/.     References 1. El Hokayem, Jimmy, Holly N. Cukier, and Derek...Interview with Darrell Kotton Four Factors that Differentiate the Stem Cell Field Sign Up to Write for the...

...can be used with or as fluorophores in several different ways. Researchers have used oligonucleotides in...were born. Aptabeacons can be created in many different ways, but the general premise is that an aptamer...relying on a lack of fluorescence for readout may render this approach less sensitive in most applications.... As a product of SELEX and FACS, Broccoli is inherently more stable in cells. The new aptamer also has...stable and can go through cycles of denaturation/renaturation. In vivo assays in which protein fluors are...Aptamers can be selected for these small ligands. References 1. Ma, Hanhui, et al. “Multicolor CRISPR labeling...Shiping, et al. “Aptamer-based biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February...

... to produce abundant, externally fertilized, transparent progeny (Figure 2). These features give it a ...individual to maintain. Also in contrast to mice, the transparent zebrafish embryo develops outside of the mother...researchers to observe changes to these tissues under different experimental conditions. Strategies for generating...sequencing approaches (Harland, 2018). Several different methods have been developed, relying on established...zebrafish—including the available genetic resources and transparency of the embryo—have proven useful in a variety...vertebratedevelopment using optogenetic approaches.     References El-Brolosy, Mohamed A., et al. "Genetic compensation...plasmid collection Read our molecular biology reference Find plasmids for your research ...

...per video abstract at 2,947 views. The journals Current Biology (2,500 views), Journal of Number Theory... Addgene Youtube Channel   After watching these trends for a few years, I decided to look at what online...examples for creating effective research videos.  Karen McKee, also known as the science videographer, is...access online journals or even use a computer.  Currently, two of the more popular biology videos are:   ...published in a recent issue of the open access journal Current Biology. Video on We Share Science: http://wesharescience.com...wesharescience.com/pin/7472 Article: http://www.cell.com/current-biology/abstract/S0960-9822(16)30873-9 Youtube:...WeShareScience or email Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric...

...Emily P. Bentley in December 2024. References and Resources References Anzalone, A. V., Gao, X. D., Podracky...(Anzalone et al., 2019). Plus, it can combine different types of edits with one another. All of this is... An overview of prime editing. Created with BioRender.com. How does it work? First, an engineered ...precise than base editing (in certain circumstances) Current base editors can perform any base-to-base conversion...indels than prime editing. But for positions that aren’t well positioned within the editing window, prime...type, and goal of the experiment can all impose different constraints and affect how you select and design...5.1-fold (though up to 45-fold) higher across different genomic sites compared to PE1 (Anzalone et al....

...research coming out of the lab.  References and resources References Hao, Y., Yan, J., Fraser, C., Jiang...affinity. This extended-open state is key to strengthening ligand binding, mediating cell adhesion and ...then used these antibodies to show the binding preferences of the αV subunit and its β partners. Of interest...antibodies and affinity reagents to study the difference in cell morphology changes induced by integrins...antibodies comes from IPI’s work with the Boston Children’s Hospital and Harvard-based lab of Tim Springer...discovered by yeast display reveal αV subunit pairing preferences with β subunits (p. 2024.01.26.577394). bioRxiv...

...disease postnatally. Now, three papers published concurrently in Science have shown CRISPR can treat a genetic... Even low-frequency editing could make a huge difference in DMD, and it’s estimated that exon skipping...respectively. Although each study used slightly different methods, they each observed the same positive ...necrosis compared to unedited animals. In grip strength, specific force, and other muscle tests, muscle...disease type C. Although the three DMD studies referenced here represent a great step forward for CRISPR...many distinct CRISPR therapies to match these different mutations. For each therapy, effectiveness and...treatments may also be closer than we might expect.   References 1. Kaiser J. CRISPR helps heal mice with muscular...

... you need. Most often, early career scientists aren’t trained to have these types of negotiations. They...all of the major equipment that you use in your current lab first. Generally, major equipment can be defined..., glassware, etc), get a simple list from your current P.I., lab manager, or administrator containing ... Do online searches, and seek advice from your current administrative services, managers, and techs’ financial... is where you can start to identify which items aren’t critical to your research success. If you can get...can still go horribly awry when several factors aren’t taken into consideration. 1. Always focus on the...for the success of your science. If the resources aren’t available, then you’re entering a win-lose situation...

... for in the future! References Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, Waldhauer...of Cas9 activity.  The field of optogenetics is renowned for enabling precise temporal and spatial control...CRISPR gene activation system, but settled on a different optimal fusion protein combination. The optimized... C-terminal (residues 714-1368, C714) halves, rendering the Cas9 non-functional when split but regaining...absence of light, Cas9 is bound by AcrIIA4 and rendered inactive. In the presence of blue light, AcrIIA4...Alexander Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically...photoactivatable transcription systems to induce neuronal differentiation. Nat Methods 14:963–966 . https://doi.org/10.1038...

...However, many genes have different functions in different cell types, at different times, and in combination...T cell responses to cancer.     References and resources References LaFleur, M. W., Lemmen, A. M., Streeter...combination with different genes. Having CRISPR systems for assessing genes in defined contexts would be quite...

... tissues are nearly transparent. As Verkhusha explained it to me, the transparency window of mammalian...tumor. They wanted to find something better. Transparency window That’s exactly what they reported doing...corals, but all of them fluoresce outside the transparency window of mammalian tissues,” Verkhusha said...near-infrared fluorescent proteins within this transparency window, so now, we could see deeper.” Adding...of the time, the answers will be there.     References: Bright and stable near-infrared fluorescent ...

...Emily P. Bentley in September 2025. References and Resources References Chen, L., Zhu, B., Ru, G., Meng, ...component of the nucleoside inosine. Created with BioRender.com. How does base editing work? Base transition... UGI = uracil DNA glycosylase. Created with BioRender.com. The BE3 system increased editing frequency...APOBEC3G displayed less predictable sequence preferences.  The Liu lab also used natural and engineered...yielded an editor with more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more...the development of prime editing, which uses a different mechanism than base editing, has allowed scientists.... A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing. Nature Biotechnology...