...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...

...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....

...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 ...

... 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 ...

...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 ...

...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...

...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...

... 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...

...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 ...

...? 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 ...

...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...

...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...

...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...

... 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 ...

... 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 ...

... 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 ...

... 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 ...

...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...

...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...

...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...