...to only four days (Paix et al., 2017). The co-CRISPR and co-conversion approaches have the advantage of...using a co-injection marker that is mainly lost in F2 animals, removing the need to segregate away co-conversion...knock-ins at other genomic loci. The first approach – co-CRISPR from Craig Mello’s lab – used inactivation...oligo-mediated knock-in of a dominant mutation, known as co-conversion (Arribere et al., 2014). In both cases...amounts of experimental manipulation and screening.  A co-selection strategy for CRISPR editing in C. elegans...approaches. Being a “lazy” geneticist, I developed a co-selection approach relying on repair of a conditional-lethal...center images depict the power of lethal mutation co-selection. Only animals rescued for the conditional...

...expect from a co-op experience.  Sarah Gordon is a rising junior at Northeastern and Addgene co-op studying... to complete a six month work experience called a co-op to receive a degree. Northeastern compiles a list...position at Addgene as a marketing and social media co-op. There are a lot of different worries that come... made the right decision to choose Addgene for my co-op, but I know it didn’t take long. It may have been... “All Addgene” meetings, when Melina, our CSO and co-founder, approached me and asked about my current...building out social media spreadsheets every day of my co-op. I had to do research on loads of different best...

...chosen field. This cooperative education experience (co-op) is what has brought me, a second year bioengineering...first saw the comprehensive job description for this co-op, I was immediately interested in applying. This...accepted that day and was excited for the start of my co-op in January. This excitement did not come without...shows how they fit into the system. Specifically as a co-op, it is encouraged to explore different teams and.... She is one of several students completing their co-op at Addgene. This is her second contribution to...

...Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists...scientists use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple...scientists have instead developed novel methods to co-express your gene along with a reporter, such as ...in the same cell. Although it is possible to drive co-expression by using a plasmid with multiple, individual...another ribosome recruitment site, thereby resulting in co-expression of two proteins from a single mRNA. IRES...efficiency. How do I get started? If you are looking to co-express your gene of interest along with a fluorescent...Lentiviral Ac5-STABLE2-neo T2A Insect   For the co-expression of multiple unique genes, you can start...

...to become a product. I spoke with Dr. Mike Koeris, co-founder and current VP of Business Development & ...from academia to a startup Mike Koeris and fellow co-founders Tim Lu and others first came up with the...skills necessary to be a succesful entrepreneur? As a co-founder of a biotech startup, Mike explained, his...self-sustainable. Within a year from this decision, Mike and his co-founders raised $6 million. The biggest challenge...Beckman Acknowledgments Thank you to Dr. Mike Koeris (Co-founder and formerly VP Operations of Sample6) for...

...more than one vector to express the gRNAs and Cas9. Co-transfections can lead to variable expression levels...expressed from a polycistronic transcript  Requires co-transfection of Cas9 Nissim et alTsai et al PTG...expressed from a polycistronic transcript Requires co-transfection of Cas9   Xie et al  Multiplexing...most of Zetsche et al’s experiments, a single vector co-expressing Cpf1 and a crRNA array were used.  With...the targeted brain region and ~75% of neurons were co-transduced with Cpf1 and GFP.  Of these neurons, ...

...Fan, Kenneth Fan, and Benjie Chen! Melina Fan is a co-founder of Addgene. Melina received her Ph.D. in ...earned a B.S in Biology from MIT.   Kenneth Fan is a co-founder of Addgene. Kenneth received his Bachelor...Technology Group in Palo Alto, CA.  Benjie Chen is a co-founder of Addgene and received his PhD in Computer...

...collection of memorabilia, even made an appearance during co-founder and CSO Melina Fan's words of welcome! Executive...contributions of each and every Addgenie, from our co-ops to our directors, we can't celebrate Addgene ...

... a graduate student at Harvard University, former Co-director of Science in the News, and member of the...received her PhD from Harvard University, is a former Co-director of Science in the News, and is a member ...

...fused the Cas enzyme to cell-penetrating peptides and co-incubate it with an “assist peptide” that facilitates...viral sgRNA targeting mCherry reporter (mChe), with co-incubation with assist peptide (AP). C) Workflow ...Workflow for Cas-RNP-PAGE editing of primary T cells, by co-incubation of Cas-RNP and assist peptide. Adapted... brains with brilliance By Hridaya Patel (Addgene co-op) Researchers in the Gradinaru lab have developed...

...is Lauren Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst intern...interview series was born. Over the six months of my co-op, I have conducted informational interviews with...Staying true to your values While going through the co-op application process, my advisor encouraged me ... project was one of the biggest takeaways from my co-op. It has been so insightful to learn about different...

...clinical development programs in biopharma. AveXis, co-founded by Kaspar, acquired by Novartis and rebranded...the mutation (Le et al., 2019). Beam Therapeutics, co-founded by Harvard Investigators and Addgene depositors...syndrome program being pursued at Vico Therapeutics, co-founded by Gail Mandel. There are numerous technical.../10.1002/1096-8628(200022)97:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge...

...editor is fused to a fast-splicing split intein and co-infected into the cell. Intein splitting removes ...1: The base editor is split into two plasmids and co-transfected into cells for reconstitution. Image ... permission. Though the lab suspected that the co-transduction by two viruses would lower editing efficiency...

... family members, and professional network such as co-workers, professors, and mentors. Ask for specific...economically disadvantaged populations. Roo is the co-founder of STEAMid.       Rose C. St Pierre is currently...help their children thrive academically. Rose is the co-founder of STEAMid.     Additional resources on the...

...double-strand break (DSBs) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are ...etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a wide variety...Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes...

... a graduate student at Harvard University, former Co-director of Science in the News, and member of the...received her PhD from Harvard University, is a former Co-director of Science in the News, and is a member ...

...genome loci, the most efficient and easiest way is to co-transfect several plasmid together, with each plasmid...clone two spacers for each locus into PX330, then co-transfect or delivery the two plasmids together into...use double nickase, you need to have four spacers co-expressed and using PX335 to express the Cas9n (nickase..., test run them and then select the best guide to co-transfect in HR donor. Alternatively, a new system... 10uM oligo stock for each well on 24-well plate, co-deliver into the cells with the Cas9/sgRNAs vector...screened? Before starting your experiment, consider co-transfecting with GFP. This allows you to sort for...puromycin resistance cassette, such as PX459 ). After you co-transfect the CRISPR/Cas system with your homologous...

... her brother Ken Fan, and her husband Benjie Chen co-founded Addgene in 2004. They worked tirelessly to...the Room or Habitat for Humanity take you and your co-workers out of the office and into a completely new...mix It’s easy to sit around and just eat with your co-workers. Don’t fall into that rut! We’ve done everything...

...screened? A4: Before starting your experiment, consider co-transfecting with GFP. This allows you to sort for...puromycin resistance cassette, such as PX459). After you co-transfect the CRISPR/Cas system with your homologous...

...field and every attendee of the meeting will be a co-author of the guide. We hope that this guide will...02139 Organizing Committee Chair: Dr. Melina Fan, Co-Founder and CSO, Addgene Organizing Committee: Dr...your participation in this meeting, you will be a co-author on this communication and you will be helping...