...discovery. CRISPR is a great example (the top requested plasmids from 2015 and 2016 were CRISPR plasmids), but...reason, the top requested plasmid of 2017 is the lentivirus envelope plasmid pMD2.G from Didier Trono’...Plasmid technologies are constantly evolving, but sometimes a technology is so useful it forever enhances...’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000 times (over 800...the broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin...ubiquitin ligase, cellular senescence, autophagy, CRISPR, and so much more (find the full, searchable list...to check out the Trono Lab website. If you have plasmids you’d like to deposit with Addgene, be sure to...

...Addgene’s CRISPR Guide Page Find CRISPR plasmids at Addgene ...GeoCas9 a try, as always, the plasmids are available from Addgene. Happy CRISPRing! References Harrington,... Read about CRISPR Methods for Bacterial Genome Engineering   Interested in using CRISPR/Cas9 RNPs? Check...Other Blog Posts on CRISPR Additional Resources on Addgene.org Brush Up on Your CRISPR Background with ...opened up the field of thermophilic bacteria to CRISPR/Cas9 genome editing. How was GeoCas9 discovered... The discovery of GeoCas9 clears the way for CRISPR/Cas9 genome editing in facultative and obligate... crispr...

...of the new plasmids, antibodies, and viral preps in our repository through our Hot Plasmids articles.  ...pooled CRISPR screens By Emily Bentley Building on their CROPseq method for optical pooled CRISPR screening...2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein (Plasmid #141386-LV [+M], negative...Huntington’s Disease, including plasmids, antibodies, viral vector preps, CRISPR tools, and more.  ...Multiplexed perturbation and decoding in pooled CRISPR screens Improved prime editor PE7 Antibodies to...developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi (Walton et al. 2024... over the original CROPseq. Find CROPseq-multi plasmids here Walton, R.T., Qin, Y., Blainey, P.C. (2024...

...received over 13,000 plasmids requests for COVID-19 related plasmids. These plasmids have reached labs in...COVID-19: CRISPR-Based Detection Tools. The CRISPR Journal 3:142–145. https://doi.org/10.1089/crispr.2020.0030...small collection of plasmids for coronavirus research, including a handful of plasmids that have been in...some highlights of the plasmids shared over the last year and how these plasmids have been used to accelerate...circulating and plasmids containing these mutations have been deposited at Addgene. Plasmids deposited from... labs also deposited plasmids for producing COVID-19 diagnostic enzymes. Plasmids in this toolkit include...These plasmids contain both untagged and tagged versions of SARS-CoV-2 ORFs and include plasmids that ...

...this CRISPR plasmid can be created in a single tube. You can also find additional SapTrap plasmids used...SapTrap is a reagent toolkit and plasmid assembly line for the creation of CRISPR targeting vectors that each...more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids 101 blog posts Resources on...Bargmann’s keynote speech and in the workshops on CRISPR techniques and new tools for conditional expression...discussed at the meeting, in addition to useful plasmids and toolkits available at Addgene.  C. elegans... development (Kyoda et al., 2012).  C. elegans plasmid tools at Addgene In addition to the wealth of open...open resources described above, there were many plasmid tools presented at the C. elegans meeting. Here...

...Silvana Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in... the CRISPR system was a large one.  The question for her then was this: Can you engineer CRISPR-Cas9 ...Two-way CRISPR screening In October, Jonathan Weissman’s lab introduced additional pooled CRISPR libraries...CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's...lab describe just such a system, which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM...tetraloop and stem-loop 2; 3. The MS2-P65-HSF1 plasmid which expresses the activation helper protein. ...earlier by the Zhang lab for GeCKO, a lentiviral CRISPR library enabling genome-scale, knockout screening...

...popular blog post on early user experiences with CRISPR alternative, NgAgo. As you might expect with any...up and running in their labs. Pooran surveyed a CRISPR google forum to get a better idea of how many people...Directed Mutagenesis by PCR & Single Base Editing with CRISPR For my editor’s choice picks this month, I’m picking... can use PCR to make small sequence changes to plasmid DNA. This an oldie but a goodie, and in my personal...experience, is a quick way to generate many mutated plasmid constructs. The second post was contributed by ...Addgene Scientist Mary Gearing and discusses a newer, CRISPR-based technique to generate C->T or G->A transitions...cells. This technique uses catalytically inactive CRISPR endonuclease (dCas9) fused to a deaminase to make...

...Background with Our CRISPR Guide Find CRISPR Plasmids for Your Research Check out Our List of gRNA Design Tools...the Ideal CRISPR Software for Your Needs Resources on Addgene.org Catch up on Your CRISPR Background... a Postdoctoral Researcher in Jacob Corn’s lab. CRISPR-Cas9 (Cas9) is an RNA-guided nuclease that targets...gene is worse than leaving the mutation alone). CRISPR delivery by nucleofection The in vivo editing experiments...validated editing reagents can be found on our plasmid pages. We encourage any labs performing genome ...genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA." Nature biotechnology...human genome engineering by controlled timing of CRISPR/Cas9 delivery." Elife 3 (2015): e04766. PubMed ...

...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'... Penn State Protein Ladder Engineered miniature CRISPR-Cas system for mammalian genome regulation and ...gene perturbation by: Gabrielle Clouse While many CRISPR-based genetic screening methods have primarily ...Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system is a cost-effective alternative... ladders. Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need... recent plasmid deposit please sign up here.      Listen to this episode of the Hot Plasmids podcast ... (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids contain...using the CRISPR-Cas9 system CHIME Article contributed by Aliyah Weinstein  Listen to the CRISPR podcast...responses in C. elegans. Find the split BoTN/B plasmids at Addgene. Liu Q et al., Neuron. 2019. PubMed...potentially problematic for imaging. Find the ARG plasmids at Addgene. For more details on acoustic reporters...

...The natural CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to ... invading phage pathogen. Over the past decade, CRISPR tools have leveraged such host-defense mechanisms...with only minimal payloads. DAP array  Although CRISPR-Cas12a, with abilities to process and release its...leading the edge of the precision gene editing. CRISPR-Cas9, the key component to enable efficient base...multiple gRNAs for nCas9-BE, or any other kinds of CRISPR tools, without compromising their strategic advantages...for potential in vivo applications or multiplex CRISPR screening.   Using tRNAs Luckily, a fun feature... state-of-art base editor, prime editor, or any CRISPR-based gene editors, for correcting mutations or...

...enChIP Plasmids? All Hodaka Fujii lab plasmids Sign up to receive email alerts when new plasmids from ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene... shopping for a wide range of plasmids. The merit of depositing plasmids to Addgene has been spectacular... molecule technologies such as TAL proteins and CRISPR became available, I was very excited because genome-editing...recently developed a retroviral enChIP system using CRISPR technology. Can you describe this technology and...retroviral enChIP system enables us to express the CRISPR complex much easier in various cells, including...user-friendly and precise. We will deposit the plasmids of these systems to Addgene in the near future...

...Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History ...transcription from the accessory plasmid (AP) encoding phage gene III. The accessory plasmid library contained all...In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted...PAM flexibility. In this system, the mutagenesis plasmid (MP6) permits quick, large-scale mutation of the...doi: 10.1038/nature26155 PubMed PMID: 29512652 Plasmids from this paper are available at Addgene.  Additional...Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed Repair...

...Addgene.org Browse our CRISPR guide pages Find bacterial CRISPR plasmids ...specific mutations in your plasmids Use REPLACER Mutagenesis to mutate your plasmid Resources on Addgene....within a user-defined genomic region. EvolvR, a CRISPR-Cas9 based targeted mutagenesis method developed... targeted mutagenesis experiments? Find EvolvR plasmids at Addgene! References Gaj, Thomas, and Pablo...Pablo Perez-Pinera. "The continuously evolving CRISPR barcoding toolbox." Genome biology 19.1 (2018): 143...PMCID: PMC6154929. Halperin, Shakked O., et al. "CRISPR-guided DNA polymerases enable diversification of...resources on the Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn...

...Get the basics about CRISPR in our CRISPR guide Browse our entire COVID-19 plasmid collection ...technologies like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID...check out this table.  All-in-one dual CRISPR/Cas12a (AIOD-CRISPR) assay – for rapid, ultrasensitive and...detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-100 copies of...Lucia et al., 2020). Detecting coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic...the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification was able to detect as ...varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources Many thanks to our guest bloggers...

...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...targeting range of CRISPRCas9 by engineering SpCas9 Article contributed by Shreya Vedantam CRISPR/Cas9 is widely...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        New toolkit...2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method to tag endogenous proteins in... the Kanemaki lab has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins ...and other tags (Yesbolatova et al., 2019) using CRISPR. This toolkit makes it easy to tag your protein...

... on Addgene.org Check out Our CRISPR Guide Pages Find CRISPR Plasmids for Your Research Find Validated...additional, unique facets of CRISPR DMD therapy. Long et al. show that AAV-CRISPR does not cause obvious germline...directions and obstacles to clinical CRISPR editing Given the success of the CRISPR-mediated exon skipping approach...To bring DMD CRISPR therapy closer to the clinic, much work is still needed. First, CRISPR delivery must... to be treated clinically with CRISPR. Many obstacles remain for CRISPR gene editing, but given the speed...Having seen CRISPR’s success in basic research, researchers are eager to apply it in a clinical setting...setting. CRISPR is often used for animal germline modification, to repair or add in disease-causing mutations...

...editing Read more about CRISPR Learn all the basics about CRISPR in our CRISPR 101 blog series Resources...following components: A plasmid in the host bacterium expressing dCas9 A plasmid in the host bacterium ...base editors for your research Read our CRISPR guide Find plasmids for your research ... Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and Beyond meeting earlier this month... of gene III to create infectious particles. A plasmid-based genetic circuit in the bacteria links the...encoding the deaminase TadA-7.10 A mutagenesis plasmid controlling the phage mutation rate Phage infection...Phage-assisted evolution of base-editing activity. A plasmid-based genetic circuit in the bacteria links the...

...you’re getting started with them. CRISPR Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening...publication. Optimizing Donor DNA for Enhanced CRISPR Genome Editing CRISPR has revolutionized how researchers make...Nathaniel Roquet from the Lu Lab at MIT developed plasmid based systems that report the order in which specific... can use CombiGEM to barcode small libraries of CRISPR gRNAs to use for screening experiments. Get the...and other types of genome edits, but, even with CRISPR, it isn’t always easy to insert a section of DNA...biophysics! Casilio: An Adaptive, Multitasking “CRISPR-OS” In this post, Albert Cheng and Mark Wanner ... plasmid...

...specifically, the Addgene-Seeding Labs plasmid grant provide plasmids to these researchers. In the third ...the Addgene-Seeding Labs Plasmid Grant Check Out Addgene-Seeding Labs Plasmid Grant Awardees in 2016 and...Resources at Addgene.org Check out our CRISPR resources and CRISPR for plants Bacterial Expression Systems...to provide scientific resources, equipment, and plasmids to researchers in resource-limited settings. More...third round of the Addgene-Seeding Labs plasmid grant we are excited to introduce the 5 awardees and their...PZA-resistance by using different approaches, including CRISPR and recombination-mediated genetic engineering ...more information on bacterial engineering using CRISPR on our blog). Her research should provide us with...