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We narrowed to 609 results for: CaS;

Showing: 201 - 250 of 609 results

  1. Another Pathway into Cells: iTOP

    Type
    Blog Post
    Published
    June 23, 2015, 8:37 p.m.
    ...the usefulness of CPPs must be evaluated on a case-by-case basis. While setting up their CPP system to ...would be compatible with CRISPR/Cas9 genome editing. Although CRISPR/Cas9 is all the rage, they had another... method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only transduction? Niels...expression. Image courtesy of Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested iTOP...expression has a long-lasting effect, such as Cre- or Cas9-mediated recombination. Both primary hESCs and immortalized...cells treated with iTOP efficiently took up both Cas9 protein and a gRNA (yes, iTOP can also be used for... indicating that iTOP may be valuable for CRISPR/Cas9 stem cell editing. The small molecule NSDB...

  2. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...To overcome this obstacle, a positive selection cassette can be incorporated, and used during the differentiation...the development of site-specific nucleases (CRISPR/Cas9, TALENs, and ZFNs). However, several challenges ...piggyBac™ transposon system can be combined with CRISPR/Cas9 to efficiently perform genetic manipulations in ...along with a site-specific nuclease (such as CRISPR/Cas9) to create a double-stranded DNA break. After DNA..., but defective for integration [4], the donor cassette is removed leaving behind no accessory transposon...recombination and Footprint-Free™ removal of the selection cassette by excision-only piggyBac™. Image courtesy of ...recombinase technology (Cre/loxP) to remove the donor cassette. While this method can be effective, a loxP site...

  3. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    Published
    April 18, 2019, 12:53 p.m.
    ...fluorescent protein to various Cas9 variants, SpCas9, NmCas9, and St1Cas9. These fusions localized to specific...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...inhibition, among others. Over the past few years, dCas9 has also become a robust visualization tool to study...University of Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal... al. “Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.” Nature...

  4. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation ...question for her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that...describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is an... three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA...system, Konermann explained, was new knowledge about Cas9’s crystal structure. “I thought maybe we could use...The activator (CRISPRa) sgRNA library uses the sunCas9 system and contains 10 sgRNAs for each transcription...levels of control in the system. First of all, the Cas9s can be turned on and off, to reversibly activate...

  5. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ... kit to CRISPR/Cas9? When would you choose one or the other? Musunuru: The CRISPR/Cas9 system is easier...system for CRISPR/Cas9 (also available from Addgene) only has two plasmids. In many cases, CRISPR may be ...makes possible, and how it compares to his CRISPR/Cas9 system. Addgene: For people unfamiliar with this...two ends and put them back together, there are occasional errors. That can be advantageous if you are trying...genes that have been implicated in disease or, in cases where particular variants or mutations have been...more efficient. You might often start with CRISPR/Cas9, but TALENs do have more flexibility in terms of...to date suggest TALENs may be cleaner than CRISPR/Cas9. Reference: Quirong Ding et al. "A TALEN genome-editing...

  6. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...CRISPR/Cas9-based systems have been developed for generating knockin mice but because CRISPR/Cas9 engineering... of a stop cassette upstream of the target gene in the targeting vector. The stop cassette is floxed, ... arms that flank the gene of interest. In either case, a positive selection marker such as an antibiotic...recombination for precise gene insertion. However, CRISPR/Cas9 systems are more useful for generating knockout ...to engineer knockout mice. ZFN, TALEN, and CRISPR/Cas9 systems can all be targeted to specific regions ...introduce multiple base pair deletions, but CRISPR/Cas9 can also introduce point mutations. While using ...so Cre-mediated recombination deletes the stop cassette, which permits expression of the target gene. ...

  7. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...Institute to provide lentiviruses for delivery of SpCas9 and Cas9 nickase for genome editing. You can also enhance...facilitated their fast adoption. However, in many cases the plasmids themselves are only the starting point...Like all things in science, getting a high titer Cas9 lentivirus prep takes a bit of time and effort that... makes it unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent...expression of your gene of interest with a variety of Cas9 activators available in lentiviral format. Finally... pathways. Find Validated gRNAs to use with your Cas9 Virus Pooled libraries now available in lentiviral...be used to infect a cell line already expressing Cas9. As John Doench (Broad Institute and member of the...

  8. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the...out the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a ...interference (CRISPRi) targets a catalytically dead Cas9 fused to a transcriptional repressor to your gene...repression of protein expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on...) are transcriptional activators fused to a dead Cas9. The system is led, via a custom gRNA, to your gene...protein levels. Looking for a DD? Check out our DD-Cas9 vector! The price of tunable control These systems...Plasmid 101: Repressible Promoters CRISPR 101: Which Cas9 Do I Choose for my Experiment Additional resources...

  9. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    Published
    Nov. 15, 2016, 3:30 p.m.
    ... Listen here! Podcast breakdown (Time : Topic) 0:00 - 2:00 : Introduction to the podcast from producers...Addgene Podcast.     Additional Resources on the Addgene Blog Learn More About Alex's Work with Cas9 Activators...In this episode of the Addgene podcast, we sit down with Alex Chavez, a Clinical Fellow in Pathology ...Engineering. Alex is the first MD PhD we've had on the podcast, has worked extensively with CRISPR, and recently...Advisory Board with a particular interest in generating Cas9 based tools to enable facile control of DNA and ...

  10. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ... had a large genetic footprint (two cassettes + a booster cassette), their expression heterogeneity required...integratable cassettes, both with doxycycline inducible promoters (Fig. 2a). The first cassette contains ...control. We also added the toxin-antitoxin stability cassette Axe-Txe so that the plasmid could be maintained... that Anabaena mARGs did not require a booster cassette, either, allowing us to reduce the genetic footprint...structural gene gvpA marked with EBFP2 and the second cassette contains the P2A-linked accessory genes gvpNJKFGWV...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow...

  11. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    Published
    Jan. 16, 2024, 6:13 p.m.
    ... CHIME works through transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral...genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9 I-CHIME Inducible knockout of a single gene...Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 L-CHIME Knocks out a single gene in defined... Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks out pairs of genes sequentially...Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME systems Proof of Concept These...Haining, W. N., & Sharpe, A. H. (2019). A CRISPR-Cas9 delivery system for in vivo screening of genes in...

  12. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced cleavage...can be used to identify Cas9/CRISPR-induced genome modifications. Occasionally, multimerization of the...through homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded break. This site-directed... the repair, the template should be resistant to Cas9 cleavage. If a plasmid contains the template, site-directed...with the function of the resulting construct. In cases where the functionally important regions span several...plasmid (or other). This is particularly relevant in cases where a large plasmid with multiple functional regions...

  13. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...side of a stop cassette, upstream of an effector gene. In cells where the stop cassette is removed during...cells will express the effector or retain the stop cassette following recombination. Image from Isaacman-Beck...effector is expressed. In cells where the stop cassette remains, the effector is not expressed and the...attP sites to drive expression of either a stop cassette or an effector gene. HDVR (2X Hepatitis Delta ... Truncating the attP site upstream of the stop cassette results in decreased efficiency of recombination...preferentially leads to gene constructs where the stop cassette remains following recombination. This strategy...

  14. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    Published
    Dec. 20, 2018, 1:15 p.m.
    ...report. A scientist working in China was using CRISPR/Cas9 in an attempt to create gene-edited babies. Several...valid critiques being offered of He’s work: CRISPR/Cas9 genome editing is not far enough along, scientifically...research published on December 13 shows us, CRISPR/Cas9 techniques are continually being refined; it is ...an embryo—even if it’s not necessarily via CRISPR/Cas9. This leaves us with the second critique. Absent...agreed-upon international moratorium on utilizing CRISPR/Cas9 in humans. This topic was to be revisited at meetings...emerging technology. We could do this for CRISPR/Cas9 – or any other germline editing technique—too. As...not “what is the translational pathway for CRISPR/Cas9 research” but whether or not we will stop shuffling...

  15. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ...sequencing in selected control and CRISPR-Cas9 injected embryos. In all cases, they did not observe off-target ... samples. Ma et al. designed plasmids containing Cas9 and two different gRNAs, and electroporated them... in editing between iPS cells and embryos make a case for conducting research in embryos should our society... decide that germline editing is permissible in cases of disease prevention. However, it is important ...alternatives are applicable in many situations. In the case of hypertrophic cardiomyopathy and other genetic... in increasing viable embryos, especially in the case of older mothers, for whom egg retrieval often results...

  16. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...single-percentage gel, you will have the option to cast your own gel. Handcast gels are significantly cheaper and generate...conditions and will cover most western blot use cases. If you’re running native (non-reduced) proteins...Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life; can alter proteins due to ...250 kDa Different sizes 4–20% gradient   Handcast or premade If you need a gradient gel, the decision...already made: gradients are extremely difficult to handcast, so unless you have an expert around, you’ll likely...hazardous chemicals, and are not always consistent. Precast gels, while expensive, reduce protocol time, can... CCD camera High initial (purchase) cost Precast gel Increased cost Save money Polyclonal...

  17. SciComm with the Experts at Science in the News Part 1

    Type
    Blog Post
    Published
    March 16, 2017, 1:06 p.m.
    ...science communication. Science in the News hosts a podcast, seminars, a blog, conferences, and more for non-scientists...the interview with Science in the News, Part 1 Podcast breakdown Introduction to Vini, Amy, and SITN ...Approach Different Audiences Listen to the SITN Podcast Many thanks to our guests, Vini Mani and Amy Gilson...of Science in the News, and member of the SITN podcast team.     Amy Gilson recently received her PhD ...Science in the News, and is a member of the SITN podcast team. You can follow Amy on twitter @aigilson. ...the audio engineer and producer for the Addgene Podcast.     Additional Resources on the Addgene Blog ...

  18. Editor's Choice, July 2016

    Type
    Blog Post
    Published
    Aug. 5, 2016, 3 p.m.
    ... when compared to the most commonly used Cas9 variant, SpCas9. These properties, combined with other benefits...Generating Mouse Models Using CRISPR/Cas9 Cpf1 Update: Comparison to Cas9 and NgAgo Special Delivery: Fluorophore... or more additional posts that deserve a peek in case you missed them. July 2016 was a fantastic month...generate mouse models.   Cpf1 Update: Comparison to Cas9 and NgAgo As you might expect given the excitement...

  19. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ... gene was disrupted and silenced by a Lox-Stop cassette, resulting in the neurological deficits observed...Restoring the MECP2 gene by excision of the Stop cassette using tamoxifen-inducible Cre not only halted ...MECP2 protein, showing mutations corrected by CRISPR/Cas9 in wild-type human iPSCs. Image modified from Le...Le et al., 2019. Gene editing via CRISPR/Cas9 and RNA-mediated correction of MECP2 mutant alleles,... Klaus Rajewsky’s group recently used the CRISPR/Cas9 system to both precisely insert the MECP2-R270X ...Joung, PhD, and Feng Zhang, PhD, is pursuing CRISPR/Cas9 technology for precise correction of Rett syndrome...Rajewsky K, Chu VT (2019) Efficient and Precise CRISPR/Cas9-Mediated MECP2 Modifications in Human-Induced Pluripotent...

  20. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...resistance cassette and you aren’t introducing an additional antibiotic resistance cassette along with...Antibiotic selection Selection is straightforward in cases where the inserted DNA includes an antibiotic resistance...plasmid backbone contains a kanamycin resistance cassette, you would plate the cells on kanamycin-containing...carrying plasmids that contain a kanamycin resistance cassette will grow. However, often times, the plasmid backbone...with your gene of interest. In these cases, antibiotic selection will select for cells that have been transformed...

  21. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    Published
    Aug. 19, 2014, 3:39 p.m.
    ...bacterial selection cassette for growth and cloning in bacteria and a mammalian selection cassette to screen for...cells, you will not need a mammalian selection cassette (e.g. 1435 pSG5L Flag HA). You can also upgrade...in an isolated, controlled environment. In most cases bacterial expression vectors are inducible (e.g....with an integrating vector expressing a selection cassette with 5′ and 3′ flanking sequences matching the...fusion proteins. Pick your promoter, selection cassette and visualization vehicle to conduct your custom...

  22. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 14, 2018, 2:36 p.m.
    ...RNA with Cas13a (C2c2) Maybe you’d like to avoid editing the genome and just stick to RNA? Cas13 may be ...Corn’s lab describe how a better understanding of Cas9 biophysics can help enhance HDR. CRISPR 101: Ribonucleoprotein...The PAM Requirement and Expanding CRISPR Beyond SpCas9 The closer your edit is to the PAM site, the higher...repair. Check out this post to see if an engineered Cas9 variant might work for your experiment. PITChing...tool to help you design, pick, and close guides for Cas9 and Cpf1. This tool supports over 250 genomes, and...

  23. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ... all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “CRISPR-OS”...Medicine described Casilio, a system that uses modular RNA binding proteins, dCas9, and a modified gRNA... the Casilio system works and how you can customize it for your experimental needs. Note: Casilio is no...

  24. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

    Type
    Blog Post
    Published
    Dec. 6, 2018, 1:34 p.m.
    ...Alyssa Cecchetelli  Listen to the C. elegans RNAi podcast segment RNA interference (RNAi) is extensively ...Eric Perkins  Listen to the TurboID and miniTurbo podcast segment A fundamental approach to studying membranes...by Izzy Mueller  Listen to the viral detection podcast segment As the primary cause for both the stomach... the absence of the target RNA sequence (in this case, norovirus RNA), the ribosome binding site is inaccessible...gene on the toehold switch to be expressed. In this case, the gene encodes the LacZ𝛼 protein. Once expressed...contributed by Michael Lemieux  Listen to the SH3 domain podcast segment The human proteome contains a breadth of...

  25. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...from the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette. It can be used to produce a ~10-fold ...and selection marker removal. CRISPR technology pdCas9-humanized (Plasmid #44246) Expression of a catalytically...catalytically inactive (dead), human codon-optimized Cas9 under the control of Murine Stem Cell retrovirus...carries CRE recombinase and a puromycin resistance cassette. No time to produce virus? Get the ready-to-use... (shRNA, miR-30-shRNA), mutation/editing (CRISPR-Cas9) or deletion (Cre), together with expression of ...

  26. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...editing technique that relies on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing...., 2019), disrupting its canonical structure and Cas9 binding (Nishimasu et al., 2014). Figure...pegRNAs that use other bases in this location bind to Cas9 with canonical sgRNA structure (right). Created ...along with your intended edit. This prevents the Cas9 nickase from re-binding and nicking the newly synthesized...Zhang, F., & Nureki, O. (2014). Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA. Cell, ...doi.org/10.1016/j.cell.2014.02.001 Yan, J., Oyler-Castrillo, P., Ravisankar, P., Ward, C. C., Levesque, S....

  27. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ... the ability to correctly assemble a lacI/lacZ cassette (designed by NEB for use in optimization of assembly...sequence for beta-galactosidase in the lacI/lacZ cassette. Additional confirmation of accurate assemblies...colonies showed a mixture of mis-assemblies and very occasional uncut or cut/re-ligated pGGA destination plasmids...12- and 24-fragment assemblies of the lacI/lacZ cassette, and illustrates how the volume of the 1 ml outgrowth...and assembly efficiency Five fragment lacI/lacZ cassette assembly was easily achievable with high levels... 12- and 24-fragment versions of the lacI/lacZ cassette were designed and synthesized. In conjunction ...

  28. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...from single or multiplex CRISPR-Cas9 double-strand DNA breaks using SpCas9. More features will be added ...Currently, ICE is configured for Cas9 fromStreptococcus pyogenes (SpCas9). ICE Score - The editing efficiency...topic page xCas9: Engineering a CRISPR Variant with Pam Flexibility RNA Editing with Cas13 and REPAIR ...

  29. Genome Engineering, CRISPR, and Research in Singapore: Interview with Wei Leong Chew

    Type
    Blog Post
    Published
    Feb. 6, 2018, 2 p.m.
    ...In today’s podcast, we sit down with Wei Leong Chew, a researcher at the Genome Institute of Singapore... Listen to the interview with Wei Leong Chew! Podcast breakdown Intro: Wei Leong Chew's career path and...the audio engineer and producer for the Addgene Podcast.      Additional Resources from the Addgene Blog...featured topic page Check out our Hot Plasmids Podcast Listen to our interview with synthetic biology ... addgene podcast...

  30. 8 Great Depositor Research Stories

    Type
    Blog Post
    Published
    Dec. 15, 2022, 2:15 p.m.
    ...Sapozhnikov tells us how to use dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation...-red, a bright red pigment reporter for plants.  Cas7-11 and PASTE: Omar Abudayyeh and Jonathan Gootenberg... worked with us to share their new CRISPR tools, Cas7-11, a low-toxicity RNA editing tool, and PASTE, ...ancestral bias in CRISPR guide library design.  dCas9-directed DNA Demethylation: Guest blogger Daniel...

  31. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ... hampered by the fact that even small Cas9 homologs like SaCas9 and their requisite gRNAs take up essentially...based system, the efficiency of gene editing is occasionally less than 1%, and virtually never exceeds 40%...et al 2017 use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template in ...: PMC3277219. 6. Ohmori, Tsukasa, et al. "CRISPR/Cas9-mediated genome editing via postnatal administration...Porteus. "CRISPR-mediated integration of large gene cassettes using AAV donor vectors." Cell reports 20.3 (2017...

  32. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

    Type
    Blog Post
    Published
    Jan. 21, 2016, 4:52 p.m.
    ...interview and happy listening! Download the podcast here! Podcast breakdown (Time : Topic) 0:00 - 1:25 : Introduction...In the third installment in our podcast series, we chat with new Addgene Board Member, Michael Koeris...the audio engineer and producer for the Addgene Podcast.     Additional Resources Visit the Sample 6 website...the Addgene Blog Check the Other Posts in Our Podcast Series Get Additional Startup Advice Learn How ...

  33. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

    Type
    Blog Post
    Published
    Sept. 9, 2016, 2:43 p.m.
    ...? While properly designed perturbations – in the case of screens using CRISPR technology, gRNAs to knockout...formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual gRNA from this single vector) ...only an gRNA, for use with cells that already have Cas9). Each library consists of approximately 3,000 gRNAs...activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology (2016). PubMed PMID: 26780180...Learn to Conduct Genome-wide Screens with CRISPR/Cas9 Get John Doench's Tips on gRNA Design Check Out ...

  34. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    Published
    May 18, 2023, 1:15 p.m.
    ...binding affinity (Janda et al., 2016; Torres & Casadevall, 2008) but…does it really do anything? Fc functions...an antigen-bound antibody and induce (or, in the case of the Fc receptors FcγRIIB1 and FcγRIIB, inhibit...system's classical complement pathway, starting a cascade of events that results in the pores being opened....2009.139 Janda, A., Bowen, A., Greenspan, N. S., & Casadevall, A. (2016). Ig Constant Region Effects on Variable...articles/10.3389/fmicb.2016.00022 Torres, M., & Casadevall, A. (2008). The immunoglobulin constant region...

  35. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    Published
    Jan. 30, 2014, 3:29 p.m.
    ...inactivates or completely removes (in the case of Gateway cloning) one cassette. Counterselection allows the scientist... Scientists introduce an antibiotic resistance cassette within the coding region of the gene they are ...is best practice not to use the same resistance cassette for the mutation and for plasmid selection. Additionally... vector typically has two separate resistance cassettes and your gene of interest is cloned into/inactivates...

  36. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    Published
    June 9, 2022, 1:15 p.m.
    ...independence, such as museums, YouTube channels and podcasts, science pub nights, and pop sci books. Most scicomm... that is widely used but has a very specific use-case (definition) in a specific field.  For example, ...also students in AP biology classes Creating a podcast about genetics using dog breeds as an example ...interested in breeding dogs who also enjoy listening to podcasts A series of Tweets breaking down a new publication...others from accessing their work. But that’s not the case. In fact, it’s really the opposite. Unlike emails...

  37. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    Published
    April 25, 2023, 1:15 p.m.
    ... the flexibility needed for multiplexing. In the case above, you can’t costain with your desired antibodies...easily manipulated into more useful tools. In the case of isotype conversion, the variable regions of a... species and/or isotype (Liddell 2013). In most cases, the resulting chimeric antibody retains the binding...properties (Morelock 1994). While in the majority of cases the differences are subtle, chimeric antibodies ...when multiplexing with chimeric antibodies. In the case above you should use an anti-rat IgG1 secondary ...

  38. Early Career Researcher Toolbox: Free Tools for Science Writing

    Type
    Blog Post
    Published
    April 28, 2021, 1:15 p.m.
    ... PhD Podcast episode 15 "Simple tricks for time management: the pomodoro technique": This podcast talks... References and Resources You Can Grad School podcast episode 2 "Organizing Literature" : I found out...out about Zotero here Dear Grad Student podcast episode 2 "Essential Grad School Resources: Citation Managers...Managers, Programming, & Productivity": This podcast talks about Mendeley, Zotero, and other citation ...

  39. Celebrate National Book Month with Addgene

    Type
    Blog Post
    Published
    Oct. 3, 2019, 12:47 p.m.
    ... carefully presented case for better more diverse science. It's a series of case studies in what happens... other works of Dr. Sacks, Musicophilia includes case studies of relevant neurological atypicalities, ...the Dark Lord rules all, the books center around a cast of endearing characters who decide to take matters...Ice Age to the 1980s, Sarum follows an intriguing cast of characters from five different families over ...

  40. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ... to help you find your desired plasmid. In many cases, you will need a screening strategy as well. In ...comes in. Let’s take a look at All_in_one_CRISPR/Cas9_LacZ, a CRISPR gRNA plasmid from Lynne Postovit’s lab..., without the need to prep your plasmid. In this case, you’d be using colony PCR to detect your insert...the other primer anneals to the backbone. In this case, if there is no insert in the plasmid, you won’t...order of nucleotides within a DNA molecule, in this case a plasmid. Sequencing is one of the surest ways ...

  41. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...selective degradation of FKBP12F36V-BRD4 using CRISPR/Cas9-mediated knock-in, and to evaluate in vivo degradation... and endogenous ENL was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7 and dTAG...and endogenous MELK was inactivated using CRISPR/Cas9 gene-editing. Featured molecules: dTAG-7, dTAG-13...engineered to express YY1-FKBP12F36V using CRISPR/Cas9-mediated knock-in. Featured molecule: dTAG-47 Additional...Knock-In and Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology Directed Repair Plasmids...

  42. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...progestin hormone therapy resulted in 76,000 fewer cases of cardiovascular disease, 126,000 fewer breast ...methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender analysis...analysis leads to discovery and innovation. One such case study looks at the effects of using all female cells...also consider the sex of the stem cells. In this case, they found that sex-matching of donor and recipient...analyze these factors during an experiment. In the case of stem cells, these factors may include cell type...

  43. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology (2016). PubMed PMID: 26780180...transcriptional activation by an engineered CRISPR-Cas9 complex." Nature (2014). PubMed PMID: 25494202. ...PMC4559080. 8. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening in human cells." Science 343.6166..."Genetic screens in human cells using the CRISPR-Cas9 system." Science 343.6166 (2014): 80-84. PubMed ...Wong, Alan SL, et al. "Multiplexed barcoded CRISPR-Cas9 screening enabled by CombiGEM." Proceedings of the...

  44. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...Yoon, S., & Kim, H. H. (2019). SpCas9 activity prediction by DeepSpCas9, a deep learning–based model with...2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase) ... and PE5 prime editing strategies) based on a DeepSpCas9 score ranking (Kim et al., 2019). PRIDICT saves...Steensel, B. (2021). Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance...

  45. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...noticing unusual cases of acute hepatitis in the pediatric population. Once ~400 cases had amassed in places...here it appears that the culprit for these unusual cases was either 1) a physiological effect of SARS-CoV...inactivation of the vector can still occur even in the case of no previous immunity existing for that adenoviral...References and resources References Bauzon, M., Castro, D., Karr, M., Hawkins, L. K., & Hermiston, T. ...

  46. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...death. The team used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed...continually produce antitoxin to avoid cell death. In the case of plasmids, toxin-antitoxin systems act as “selfish...for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct repeats which can be ... selects for those cells that have removed the cassette through a second recombination event between the...

  47. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...inserted the deaminase into the C-terminal domain of Cas9, which narrowed the editing window and reduced A.... gGBE consists only of engineered MPG fused to nCas9. First, MPG removes the G base from DNA to create...site. Like in most other base editor designs, the Cas9 nicks the non-edited strand to encourage the cell... using the edited strand as a template — in this case, encouraging it to incorporate a new base across...2024; Ye et al., 2024; Yi et al., 2024). In each case, a UNG enzyme was modified to recognize T and/or...

  48. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...Blainey’s lab has developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi...Anzalone et al., 2019, including modifications to the Cas9 or reverse transcriptase enzymes, inhibition of ... cell types at different genomic loci — in some cases, over 20-fold higher editing than PEmax. The authors...end of the pegRNA, which is not well protected by Cas9. This is similar to the proposed mechanism for engineered...of applications, including CRISPR activation by dCas9-VPR, prime editing, and delivery of disease-relevant...

  49. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...fluorophore-conjugated antibody commercially available. In other cases, fluorophore-conjugated antibodies may exist, but...may want to look at other methods. In the above cases, a good alternative is using a two-protein detection... antibodies that block surface proteins. In that case, you would want to look at negative selection: labeling...+ T cells, B cells, and dendritic cells. In this case, we simply need to label the lineage markers of ... a separate fix/perm cytosolic procedure in that case.     Figure 3: For intracellular labeling...

  50. Imaging Tools and Following a Passion for Basic Biology: Interview with Joachim Goεdhart

    Type
    Blog Post
    Published
    March 9, 2018, 2 p.m.
    ... Today on the Addgene Podcast, we have another interview conducted by European Outreach Scientist Benoit... Listen to the interview with Joachim Goedhart Podcast breakdown Intro Chapter 1 - How Joachim ended up...the audio engineer and producer for the Addgene Podcast.      Additional Resources from the Addgene Blog... addgene podcast...
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