We narrowed to 702 results for: abo.1

...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome...experiments, therefore, you often must make a choice about what is your top priority: location or efficiency...These Cas proteins have everything we know and love about Cas9, but with broader PAM compatibility. This alternative...

...initiating cleavage of the transcript.   Figure 1: A predicted conserved secondary structure and sequence...prokaryotic terminators and deposited several with Addgene (1). Prokaryotic polyadenylation Although mostly thought...constitutive RNA transport element lead to enhanced HIV-1 Gag expression in a context-dependent manner. Nucleic...additional helper sequences (2-3). As alluded to above, termination and polyadenylation of Polymerase II...how the Promoter Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids ...

...backbones for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids ...years of plasmid sharing  Interested in more facts about Addgene's 10 years of plasmid sharing? Check out...

...with a membrane-impermeable biotin substrate (Figure 1). Biotinylated proteins were isolated, identified ...transporters, ion channels, GPCRs, and more.  Figure 1: Workflow for cell-surface protein labeling and identification...564–571. https://doi.org/10.1038/s41586-022-05682-1 Zhang, Y., et al. (2025). Probing condensate microenvironments... and synthetic biology. Nature Communications, 15(1), 6563. https://doi.org/10.1038/s41467-024-50767-2...models, showing that this transfer increases the metabolic and metastatic capacity of cancer cells. This ...macromolecules to serve cellular functions. Along with collaborators, the Denes Hnisz Lab previously discovered that...models By Brian O'Neill The Nadav Ahituv Lab and collaborators have found a way to make adipocytes fight cancer...

...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central...can be used in the repressible manner described above, as well as the inducible manner detailed in our...necessary to have all 3 of the different binary systems above if they all behave very similarly?” The simple answer...

... for less money. The key steps to colony PCR are: 1) design primers to detect the presence of your insert...primers. There are 3 strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers...previously described. When using insert-specific primers (1), positive clones (+) will give a band, while a negative...SLIC cloning Learn about Gibson Assembly Additional Resources on Addgene.org Read about Choosing a Molecular...this type of primer does not provide information about the orientation of the insert. Orientation-specific...Orientation-specific primers: If you need information about insert orientation, then you might consider designing orientation-specific...insert Backbone-Specific Provides information about the size of the insert and if it's in the plasmid...

...which ranges between 0 and 1) so that they can be used in the rank equation above. Cut-to-insert distance...between 0 and 1) (shown in the graph below) so that it can be used in the rank equation above. Generally,...This gives each guide a score ranging between 0 and 1. The cutting frequency determination (CFD score) is...all the current code on GitHub, ask us questions about it, or send us a pull request for improving it. ...splits up the guide sequence.) If neither of the above are options, the algorithm silently mutates codons...Addgene blog Read our CRISPR 101 blog posts Learn about some considerations when designing gRNAs Find other...

...homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in the PITCh vector ~20...editing possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted ...CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from ...If you follow CRISPR research, you know all about using non-homologous end-joining (NHEJ) to make deletions...plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for complicated...more predictable than NHEJ. As seen in the figure above, short (5-25 bp) regions of homology flanking a ...Directed Repair and Non-homologous End Joining Read About Other Genome Engineering Tools like Sleeping Beauty...

...expected in another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...application and available resources.     Figure 1: Table summarizing the IWGAV five proposed antibody...like immunohistochemistry on human tissues, when (1) other strategies may not be feasible and (2) large-scale...validation: A view from the mountains. New Biotechnol 45:1–8. https://doi.org/10.1016/j.nbt.2018.08.002  Uhlen...may not be the most accessible option for many laboratories. Additionally, it can be difficult to distinguish...the captured proteins off for mass spec analysis (above). When the results come back it shows that the majority...our Selecting an Antibody blog post! Learn more about Antibodies and their many applications in our Antibodies...

... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...the hood. A 48-well plate is also prepared with about 200 ul of culture medium containing serum in each...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose...with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris...

...cell with the images from the Allen Cell Explorer (1). When studying the function of a protein or its role...various subcellular structures. Please see Figure 1 for commonly used markers for major mammalian organelles...Subcellular Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000....the context of the cell. However, much knowledge about functionality can be gained by understanding the...subcellular marker proteins, it’s possible to get an idea about the subcellular structures your protein resides ...with. These “colocalization” studies give insights about the proximity of two proteins within the same subcellular...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...

...some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...grown at 30 °C instead of 37 °C. If you’re unsure about the integrity of your virus, perform a diagnostic...Additional Resources on the Addgene Blog Learn All about Adenovirus Associated Virus New to Virus? Check ...Check out Our Primer for Beginners! Learn All about Lentiviral Vectors Additional Resources on the Addgene.org...

...expression. Brown et al., Genesis 2014 Jan;52(1):39-48. APEX2 for proteomic mapping and electron...engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing the set of... of human mitogen-activated ERK activating kinase 1 (MEK1) with either one or two phosphoserine residues...comprised of three sets of cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly... vectors are directionally assembled into a Level 1 vector creating a single transcriptional unit (Ex:...coding region, and terminator). Next, up to six Level 1 modules can be similarly assembled into a Level 2 ...vectors to allow for additional iterations of Level 1 assembly if necessary. Combining multiple Level 2 ...

...active for greater than 50 generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters...on a non-selective plate. Most bacteria tested had 1 transfer in every ~100 to 1,000 colonies, but some...including E. faecalis and P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko...

...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production... widely available, so you’ll only have to worry about cloning your transfer plasmid. The transfer plasmids...typical insect cell preparations, which we talk about below. Processing The viral supernatant is typically...available, so the amount of cloning required is about the same between insect and mammalian systems. Baculovirus...dead cells and cellular debris. While adding time (about three to four days), this second round provides ... can skip the first round of production, saving about a week of work. An important note is that this is...

...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...possible, do at least two full length practices (~1 h) and get feedback. It’s also helpful to practice...a single grant. Either way, think realistically about the time and resources required to feasibly accomplish...interrupted frequently by committee members. As mentioned above, plan to give your 10 minute overview and address... list of common questions, check out this eBook about applying for faculty positions. You need to practice...program. You should be proud of it and be excited about getting the opportunity to realize your vision. ...blog posts Resources on Addgene.org Learn more about careers at Addgene Watch our career videos ...

...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6... an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as a tool for...PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered, check out ...Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous system (...

...with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control over... transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny, J. R. (2013) ‘A robust...transformation’, Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID:... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...

...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike...Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; ...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...is multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...Check out our blog post on AAV serotypes for more about the tissue tropism of these and other AAVs. ...β-secretase; AF = AMPA.FingR; FingR = fibronectin intrabody generated with mRNA display. Modified from Rivera...