... as their orientation relative to the gRNA (5’ or 3’). In some cases, there is no requirement, which is... spCas9 isn’t always the winner!).   Fig. 3 – Comparison of different Cas enzymes using CasPEDIA...you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly has this information...

...very little bit about hiring when we need to do it quickly. Hiring is hard but as a 3 year Boston Globe...like The Atlantic. We’ve been in documentaries in 3 languages (Japanese, German, and English) and even...keep spreading the word about materials sharing, but we don’t have to worry about getting support from depositors...of course always being vigilant about resources, I’m not worried about the stewardship of this in the ...Addgene is about half of what is standard in most sectors. The Addgenies support a high collaboration, high...on this. Addgene’s success is about the Addgenies. I never worry about their dedication, excellent intentions...more about Addgene Resources on the Addgene website Find plasmids for your research  Learn about our ...

...microenvironments made of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The...microenvironment’s physiology is deeply connected to these 3 stages and contains multiple cell types: fibroblasts... related blog posts Learn about our cancer collection pages Read more about the tumor microenvironment...the tumor. How do microenvironment enzymes and metabolic reprograming influence tumors development? Being... vice versa, let’s take a look at cancer cell metabolism. Cancer cells promote oxidative stress leading...of cells affect tumor progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential...particularly interested in science journalism and writing about biomedical issues. Follow her on Twitter @BrbaraPinho6...

...way is not necessarily easy, so we came up with a 3-part strategy: Train: Nearly every new Addgene employee...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing 8...differently based on their audience. How you talk about your experiments to your PI, your colleagues, or...have cross-team projects within your own lab, collaborations with other scientists within and outside your...not happen in the lab, they have parallels to collaborative lab work. Corralling context and junking jargon...difficult to know what kind of plate they were talking about during the meetings. And “plate” certainly wasn’... a lot on context. New people in the lab or collaborators will not have the context you do, which means...

...they observed that 41/42 embryos were non-mosaic at 3 days post fertilization. Since Ma et al. designed ...proper development. Ma et al.’s experiments showed a 3% mosaic rate, and this rate would need to be much,...August 2nd in Nature gives us even more to talk about, as it represents another step towards CRISPR germline...impossible with current tools, thus quieting some fears about the creation of “designer babies.” The differences...of individuals with varied backgrounds. Talking about this study with non-biologist friends Like me, you... Blog Check out Our New CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes...

...workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single alpha helix) and target...optimized PASSIGE plasmids here!     Figure 3: Schematic of the PASSIGE strategy. First, twin prime.... When the prime editor releases the DNA, the two 3’ flaps containing the edited sequence anneal with ...transcriptional silencing (Nuñez et al., 2021). Now, in a collaboration with the Vallabh/Minikel Lab, the Weissman Lab...

...manuscript review/submission process at JEI. Chapter 3 - JEI target audience and their current readership...working in a lab for the first time, and thinking about the future. Many thanks to our guests,Olivia Ho-Shing...resources Use our Plasmids 101 eBook to teach students about basic plasmid biology Use or CRISPR 101 eBook to...

...a truncated version of the original Tol2 and has ~3-fold increase in transposition activity  Tol2 transposon...the QCascade DNA-targeting module with the crRNA, 3) a transposase complex made up by three transposase... up to 10 kB in size in bacteria.   Figure 3: INTEGRATE system for RNA guided transposon insertion...Addgene blog Learn about the INTEGRATE system for bacterial genome engineering  Read about the Sleeping Beauty...DNA transposons are most commonly used in the laboratory for genome manipulation. When transposons are...Applications of transposons Now that you’ve learned about some of the popular transposon systems, let’s take...2020) Jump around: transposons in and out of the laboratory. F1000Res 9:135. https://doi.org/10.12688/f1000research...

...help ensure ensure equal distribution. Lab tip #3: Barcode everything Use barcodes and scanners to keep...locate materials quickly and easily. Learn more about using barcodes. Check out our “Lab Tip” filter...

...pulse of 500 nm light (turn on psRaf1) followed by 3 seconds of 400 nm light (turn off psRaf1). After this...is responsible for this MEK1/2 negative feedback. 3. Studying signaling pathways in vivo Expressing photoswitchable...Additional Resources on the Addgene Blog Learn More about Photoactivatable Fluorescent Proteins Control Protein...

...produce yellow, cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created...Find the plasmids from this publication at Addgene. 3. Ma, Hanhui, et al. “Multiplexed labeling of genomic...Additional Resources on the Addgene Blog Read More about Fluorescent Protein-Based Tools Learn Some of the...

...proposal successful. Typical proposals consist of 3 to 5 aims, with secondary milestones to meet along...application to see how funds were allocated. Step 3: Get feedback from your peers and advisors Once a...first. Additionally, reach out to peers and collaborators who have succeeded at grant writing to see what...

...the stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; Pattanayak et al. 2013)...every CRISPR experiment. Off-target events due to >3 bp mismatches or that are sequence-independent are...PMID: 25566793. PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening...et al. 2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...unbiased sequencing may not be worth the cost and labor when single clones are selected. Conversely, translational... Hough is a Biochemistry PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute...

... perspective on the field. 3. Highlight your network and your collaborators It’s a good idea to move around...before submitting - handing them your application 3 days before the deadline isn’t going to be much use...introduction to my area, a line about the research I will be doing, and a line about what I hope to discover....themselves many times and will have useful advice about what reviewers in your area are looking for. Remember...One way to add originality to your work is to collaborate with specialists in different fields that can...outcomes can include poster presentations, talks about your work, or even the successful use of your preliminary...quality application. Even following this advice, only about 50% of my grant applications have been successful...

... to show people where the lab magic is happening. 3) Volunteer – Step into science education Do you aim...-to-day business with popular work experiences of 3-10 days. And the Society of Experimental Biology offers...Sainsbury Laboratory. She is passionate about science communications and loves talking about science, ...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...break into science communication? Everybody goes about this in their own way. The career paths into science...Society gives training in media skills, writing about your research and residential communication. If ...

...materials required for each [1, 2]. This protocol [3] provides, in exquisite detail, the various steps ...doi: 10.1038/nprot.2011.413. PubMed PMID: 22157972. 3. Britt JP, et al. Use of channelrhodopsin for activation...distance so ideal implant placement should be about 0.4-0.5mm above the site of your stimulation. The scatter...scatters a great deal once it enters tissue, placement above the site of stimulation guarantees that the majority...instead). Laser stimulation         In the video above, Channelrhodopsin 2 (Chr2) has been expressed in...These lasers are relatively cheap and will run you about $2-3,000/laser. However, an important caveat is ...output of the laser must be controlled. As discussed above, each opsin variant has slightly different activation...

... to have either salaries raised or hours tracked, 3% of postdocs were at institutions focused only on ...Previous Article: Changing Labor Laws Bring Increased Postdoc Wages Read about the Advocating for Science... of $47,476 per year, under updates to the Fair Labor Standards Act (FLSA). This was due to the threshold...postdocs - and in some cases, faculty - have found out about what their institution is doing from Future of Research...Science Symposium and Workshop Learn about a Variety of Science Careers Resources on Addgene.org Find Career...

...PMID: 27175853. Pubmed Central PMCID: PMC4866822. 3. McDowell, Gary S., et al. "Shaping the Future of ...perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central PMCID...purposes of labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy...passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which mandates that all workers...event of note is a ruling in August by the National Labor Relations Board to allow graduate students who work...

...expression (Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation...work well, and up to 10 base pairs away (especially 3’ of PAM) might work too.  High dCas9 and gRNA expression...Daniel M. Sapozhnikov, a graduate student in the laboratory of Dr. Moshe Szyf at McGill University. Some ...expression, confounding any potential conclusions about the role of methylation (Hrit et al. 2018, Kaas ...Daniel M. Sapozhnikov is a graduate student in the laboratory of Dr. Moshe Szyf at McGill University, working...

...knockout, activation, repression, and barcoding.  3) Are you trying to target every gene in the genome...sequencing. Representation - Most libraries contain 3-6 gRNAs per target gene, and maintaining the distribution...preparation from Addgene, you can skip the steps above! Cells are then transduced with lentivirus containing...practices, CRISPR libraries can help you learn a lot about your phenotype of interest. For more detailed information...Libraries” (McDade et al., 2016). To learn a bit about how CRISPR/Cas9 can be used in other types of experiments...

...position at the Genome Institute of Singapore. Chapter 3: Biology research and resources in Singapore. Chapter...getting a lab up and running, and learn a little bit about what it was like for Wei Leong to work in George...

...plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually encodes...vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require some initial optimization...available systems. Producing lentivirus: it's all about the T's (transfection and titers) To produce lentivirus... optimize transfection conditions. As mentioned above, plasmid ratios can be adjusted to enhance titer...

...biological study without her or her family's knowledge. 3. Fossil Rock Anthem One of Tom's earliest and most...Science with Tom. In this podcast, we learn more about Tom and pick his brain for advice on how to dive... Rap of Henrietta Lacks Students in Oakland rap about Henrietta Lacks - the African American woman whose...Please Don't Kill My Hive Beautifully filmed video about colony collapse disorder.     5. Plasmid Rap ...science standards and communicate science. Learn more about Tom on his youtube channel and at sciencewithtom.com...

...passion and pay attention to the details. Chapter 3 - Answering interesting biological questions using...of our educational resources. Listen to hear all about Professor Goedhart, his lab, and some of the tools...

...degradation of endogenous proteins (notably, BRD2/3/4, CDK9, TRIM24, FLT3, BTK, and ALK) by linking small...with dTAG-13 at doses as high as 20 µM (see Figures 3 and 5 in Nabet et al., Nature Chemical Biology for...degrade any protein. We are particularly excited about the dTAG system as we can use it to evaluate the...

...targeted sequence at the locus of interest. SpCas9’s 3’ NGG PAM occurs frequently in GC-rich genomes, but...100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent editing...Blog Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed...editing specificity. We’re excited to learn more about xCas9 - here’s what we know so far! Why do we ...point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’s a PAM...in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG PAM. When...are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9 is going to be flat...

... of the optical fiber into the region of interest 3) Behavioral experimentation I’ll discuss some of the...thoroughly elsewhere (here and here for instance) [2], [3]. Karl Deisseroth of Stanford University , one of ...10.1146/annurev-neuro-061010-113817. PMID: 21692661.  3. Bernstein JG, Boyden ES. Optogenetic tools for analyzing... electrophysiology data too but that’s what collaborators are for). For example, if you study neural pathways...intended brain region and/or cell type (I’ll talk more about this in detail later). But what are the different...of your opsin, but keep you eyes open for posts about the next steps in the coming weeks. Expression of...Addgene. See the Deisseroth lab homepage for details about types of constructs available, instructions for ...

...DNA primers that bind upstream (5’) and downstream (3’) of the DNA region of interest. Primers are crucial... the dNTPs.  Let’s jump into the specifics of the 3 basic steps of a PCR reaction:  Denaturation- To...If you have ever worked in a molecular biology laboratory you have likely done a polymerase chain reaction...uses ddPCR for AAV titrating. You can learn all about ddPCR for AAV titration and find helpful tips and...method that can help save time and effort in the laboratory.  There are two main categories of multiplex ...use a single pair of primers, like those stated above. Multiplex PCR is also used in high throughput SNP...

...ensure nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep...Journal of applied phycology27.6 (2015): 2321-2332. 3. Contamination, Ileana S., John P. S., Basic Methods...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

... the lysosome where they are finally degraded (2, 3). Once you've created your fusion protein, it's important...2000. The Cell: A Molecular Approach. 2nd Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway...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...

...Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing by a DNA-guided...success of CRISPR-Cas9 across multiple model organisms (3) lies in the versatility of its delivery methods. ...technology. To dig more into what researchers felt about NgAgo, I conducted a survey and asked researchers...successful, and the second section asked basic questions about the NgAgo protocols used by respondents (transfection...PMC4343198. Resources on the Addgene Blog Learn More about NgAgo Enhance CRISPR Targeting Specificity with ...

... The collaborative nature of working in industry and how delegating can empower people Chapter 3: How ...therapeutics at Novartis - listen to learn more about Crystal’s exciting career path, to gain insight ...metrics for success. The importance of thinking about the big picture and not just the next experiment...resources in our Mentoring for Scientists Series Learn about Turning Your PhD into a Career with Joanne Kamens...

...binds EQKLISEEDL (pronounced “equi-kli-seed-le”). (3) Tag all the things!   Myc1-9E10 and the Myc-...mapping functional domains of Drosophila hsp 70. Embo J 3:3087–3093. https://doi.org/10.1002/j.1460-2075.1984...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take...times of the myc tag.  Origins of protein tags For about as long as the disciplines have existed, cell and...examples of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable ...

...which is achieved by deleting a large portion of the 3'LTR in the transfer plasmid. The deletion prevents...number of HIV genes required for virus production (3 or 4). Third-generation vectors contain gag, pol, ...Plasmids 101 Learn about genome-wide screening with CRISPR and Lentivirus Read about the ClonTracr Library... the proper cell line is used; several common laboratory cell lines including 293 were immortalized with...

... U3; Unique 3'; region at the 3' end of viral genomic RNA (but found at both the 5' and 3' ends of the...virus production are split across multiple plasmids (3 for 2nd-generation systems, 4 for 3rd-generation systems...incompetent and may contain an additional deletion in the 3'LTR, rendering the virus “self-inactivating” (SIN)... VSV-G Safety Safe. Replication incompetent: Uses 3 separate plasmids encoding various HIV genes. Safer...incompetent and always SIN: Uses 4 plasmids instead of 3 and eliminates the requirement for Tat. LTR Viral ...is outlined in the simple schematic to the right. 3-4 plasmids are transfected into A293T cells: one transfer...SIN) vectors. These vectors have a deletion in the 3'LTR of the viral genome that is transferred into the...

...it easier to compare kits at a quick glance, Table 3 below contains details regarding kits from a variety...compare kits before deciding which one to buy. Table 3: Comparison of popular plasmid purification kit options...of some commonly used kits, please note that Table 3 is certainly not comprehensive — there are many other...columns we’ve been talking about!) are a significant source of waste in laboratories. To help combat this, ...dependent on many different factors as we detailed above, so be sure to pick what kit works best for you!...

... U3; Unique 3'; region at the 3' end of viral genomic RNA (but found at both the 5' and 3' ends of the...SIN γ-retrovirus lack most of the U3 region of the 3’ LTR and are primarily used in gene therapy research...transcription. R; Repeat region found within both the 5' and 3' LTRs of retro/lentiviral plasmids. U5; Unique 5';...the viral genomic RNA (but found at both the 5' and 3' ends of the provirus). 5' LTR in cis Acts as an RNA...proceeds through U5 and the rest of the provirus. 3' LTR in cis Terminates transcription started by 5'...packaging methods. The three components described above (envelope, packaging, and transfer) are supplied... Return to Top Frequently Asked Questions (FAQ) about γ-Retroviral Plasmids How do engineered γ-retroviruses...

...be controlled by blue light (Bubeck et al., 2018). 3. Reduce cytotoxicity of CRISPR-Cas editing in tissues...Disabling Cas9 by an anti-CRISPR DNA mimic. Sci Adv 3:e1701620 . https://doi.org/10.1126/sciadv.1701620 ...Bondy-Denomy et al., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered...Cas9 Variant for Your Next Experiment Read more about Optogenetic Control of CRISPR   Resources on Addgene.org...

...Ras pathway. The term “Ras” collectively refers to 3 proteins (H-Ras, K-Ras, and N-Ras) encoded by a family...cell 25.3 (2014): 272-281. PubMed PMID: 24651010. 3. Ledford, Heidi. "The ras renaissance." Nature 520.7547... cancer are yielding promising results in the laboratory, and Addgene is accelerating this research by...within that pathway that are of interest to their laboratory. A single mouse click on a gene of interest will...

...encapsulating the protein of interest (Figure 3).   Figure 3: Schematic representation of the MPyV system...acid, suggesting that MPyV system can improve metabolic pathway yields by encapsulating a single enzyme...

...  Chromoproteins for allelic exchange Figure 3: pTOX plasmids expressing amilCP and tsPurple genes...genetic modification integrates into the genome (Figure 3). Here, the chromoprotein is expressed and gives the... Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid ...

... Consulting firms deploy case teams, usually with 3-6 consultants, who work on analyzing the situation...the client to prioritize investments over the next 3, 5, or 10 years. Complicating these challenges is ...company to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...and why you might enjoy doing it. I’ll also talk about the skills you’ll need to do well as a healthcare...often work with such teams to help make decisions about R&D investments in either new internal efforts, ..., or through acquisitions of or collaborations with other companies. Medical Affairs groups work on building...building awareness among physicians and scientists about clinical research, drugs, the diseases they can ...

...into Stage 3 or Stage 4 cancer. These later stages of a cancer diagnosis indicate, for Stage 3, that the...common cancers like lung (about 224,000 diagnosed this year) or breast cancer (about 246,000 diagnosed this...Mesothelioma Cancer Alliance. We have learned much about the causes of cancer and the different avenues that... asbestos exposure to the time of diagnosis) is about 35 years, but it usually ranges from 20-50 years... an orphan cancer; it is rare disease with only about 3,000 new mesothelioma patients diagnosed every ...from mesothelioma several years ago – recently collaborated with Senator Barbara Boxer to introduce the ...Additional Resources on the Addgene Blog Find out More About Cancer and Addgene's Cancer Collection Pages Learn...

...EQR variant 3' NGAG SpCas9 VQR variant 3' NGAN or NGNG xCas9 3' NG, GAA, or GAT SpCas9-NG 3' NG Staphylococcus...SP); SpCas9 3' NGG SpCas9 D1135E variant 3' NGG (reduced NAG binding) SpCas9 VRER variant 3' NGCG SpCas9...jejuni (CJ) 3' NNNNRYAC Neisseria meningitidis (NM) 3' NNNNGATT Streptococcus thermophilus (ST) 3' NNAGAAW...characterized *In the table above, 3' and 5' indicate on which end of targeted sequence the PAM is located... DNA target, the seed sequence (8–10 bases at the 3′ end of the gRNA targeting sequence) will begin to...gRNA will continue to anneal to the target DNA in a 3′ to 5′ direction. The location of any potential mismatches...matters. Mismatches between the target sequence in the 3′ seed sequence inhibit target cleavage, while mismatches...

... dangle the carrot (or more popularly- cookies). 3. Do something outside of your ordinary environment...Utilize your office space too Balance point number 3 with doing fun things within your office. This way...early employees spent this remaining time by elaborately pranking each other, dressing in costume, and...frequently ask the question, “What’s your favorite part about working at Addgene?” Most Addgenies reply with an... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...rose-colored-glasses-esque picture of how everyone is kind and collaborative and all we do is have fun all day long, whistling...megaphone, or your company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring...

...the organization(s) you serve time and resources. 3. Get a mentor, find your people Find someone who is...GPCI evaluates a candidate's expertise (a minimum of 3 years experience) and experience through the candidate...on becoming grant writers. When we asked others about how they became grant professionals, mid- to late-career...late-career grant professionals have similar stories about a time when their passion for an organization or... logic models, established partnerships and collaborations, and commitments to and proof of compliance...grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers... skills & passion  Skills such as research, collaboration across multiple disciplines, writing, project...

... for an extended period of time (>10 passages or >3 months). Other times cell lines are unavailable from...State University of New York Press, Albany (1986). 3. Masters, John R. "HeLa cells 50 years on: the good...Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about the Dangers...Misidentified or cross-contaminated cells have been known about since 1966 when Stanley Gartler reported that most...there is a loss of reputation that can impede collaborations, especially with industry. Today we also have...The LGC Group also provides standards and has laboratories to carry our independent quality control for...culture contamination." Archives of pathology & laboratory medicine 133.9 (2009): 1463-1467. PubMed PMID...

...interfere with it in a BSL-2 lab instead of a BSL-3 lab. Jesse Bloom’s lab published a protocol and reagents... CRISPR-Based Detection Tools. The CRISPR Journal 3:142–145. https://doi.org/10.1089/crispr.2020.0030 ...different SARS-CoV-2 spike variants. You can learn more about their research in this article. To see other plasmids... optimized. Browse the collection or learn more about it on the blog. COVID-19 research is happening fast...through plasmid sharing and more. As you’ve read above, some of these plasmids have been in the repository...Additional resources on the Addgene blog Learn more about mapping the SARS-CoV-2 - human protein-protein interaction... interaction Find tips for remote collaborations Browse all COVID-19 blog posts Resources on Addgene....

...enhancers 3. CRISPR out the enhancers from step 2. See which ESCs die or differentiate Step 3 reveals which...republished here with permission. Quilt is a collaborative database for genomics. In this article, Quilt...tri_methylation In the next section we’ll apply the above formula to a real-world experiment. We’ll start ...discovery and therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section...Aneesh Karve is CTO at Quilt Data. Quilt is a collaborative database. Aneesh's research interests span machine...on Your CRISPR Background with CRISPR 101 Learn about CRISPR Delivery Methods Validate Your Genome Edit...

...type and the specific genes cloned into the vector.3  2A Peptides To overcome some of the disadvantages...et al. (J Virol. 1988 Aug;62(8):2636-43.) PubMed. 3. Highly Efficient Multicistronic Lentiviral Vectorswith...genes above with your gene or genes of interest. Some of the plasmids listed in the table above (and their... an internal region of the mRNA. In the figure above, you can see that the IRES element acts as another... first discovered in picornaviruses, are short (about 20 amino acids) and produce equimolar levels of ...