We narrowed to 932 results for: Gene

...editing single genes is very useful, sometimes, you need to modify multiple genes to generate a cellular ...Nicole Gaudelli, the Associate Director and Head of Gene Editing Technology at Beam Therapeutics, used a ...In addition to stem cells the authors targeted 6 genes in primary human T cells using 8 selected ABE8s ...the reduction in protein expression of these six genes. All ABE8s edited more efficiently than ABE7.10 ... was the winner. It could simultaneously edit 3 genes (98.1%, 98.3%, or 98.6% efficiency, respectively... eighth generation of adenine base editors, called ABE8 (Gaudelli et al., 2020). The generated ABE8s needed...replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM...

...limits the delivery of larger transgenes, such as base editors which have a gene size of ~5.2 kb. To overcome...Article contributed by Jennifer Tsang AAVs are a great gene therapy delivery mechanism, but their small packaging...dosage comparable to those currently used in human gene therapy trials. They further tested the base editors...expression levels from different promoters, and inhibited gene expression using CRISPRi. The kit functions in the...that increases temporal and spatial precision of gene expression and reduces toxicity as compared to previous...fluorescent genetically encoded voltage indicator Article contributed by Shreya Vedantam Genetically encoded...splitting removes any exogenous sequences and regenerates a native peptide bond at the split site. ...

...NHEJ to generate translational reading frame-altering indels for “functional knockout” of a gene of interest...) validate successful mutation of your favourite gene, there are several different methods available. ... for complete functional knockout of your target gene. Mismatch nuclease assays are completely “blind”...U., Hauber, I. & Fehse, B. Digital PCR to assess gene-editing frequencies (GEF-dPCR) mediated by designer...et al. Digital detection of endonuclease mediated gene disruption in the HIV provirus. Sci. Rep. 6, 20064...validation, such as this "CRISPR 101" post. Next-generation sequencing technologies are the gold standard.... 6. Fu, Y. et al. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells...

... et al. Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in...Well, they are essentially evolutionarily optimized gene delivery devices. Their survival depends on their...Altogether, AAV has many excellent features for human gene therapy and is currently being used in clinical ...in the New Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI: 10.1016/j.gendis.2017.04.001...v13102073 Resources on Addgene.org Adeno-associated Virus (AAV) Guide Addgene’s viral plasmids and resources...resources Addgene’s viral services: AAV Resources on the Addgene blog Viral Vectors 101: Viral Vector...due to their propensity to integrate into proto-oncogenes and disrupt their expression (Robinson, 1982)....

...Horizontal gene transfer (HGT) is the movement of genetic material between organisms. It plays a key ... comic, when a bacterium uptakes a plasmid, the genes on the plasmid can give them different phenotypes...this plasmid encodes a green fluorescent protein gene which makes the bacterium fluoresce green. Comic...used to create both transient cell lines, where a gene of interest is expressed but not integrated into...Conjugation was the first extensively studied method of gene transfer and was discovered in 1946 by Joshua Lederberg...uptake of genetic material from the environment by bacterial cells. In nature, this genetic material often...the donor can transfer genetic material to the recipient bacterium. The genetic material transferred is...

...stem cells By Jennifer Tsang CRISPR gene interference (CRISPRi) and gene activation (CRISPRa) are great ways...the plasmids at Addgene! A new tool for synthetic biology uses CRISPRi to control gene expression. This...nucleus and modulate transcription of the mCitrine gene. mCherry is constitutively expressed. Cells are ...selectivity and found that the best motif was from the Lck gene. For marking microtubules of mammalian cells, the... ways to modulate the expression of nearly any gene via specific guide RNA (gRNA) sequences. But these...-IgG1-Fc, Addgene #140700) pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) ...Find these AAVs at Addgene New controls, biosensor, chemogenetic and optogenetic AAV for expression ...

...expression time. 2. Timer functionality with a single gene Although it is possible to make a timer system composed...cargoes to FP timer mk-GO (Tsuboi et al.) Monitor gene expression in the developing pancreas by placing...FP timer DsRed-E5 under the control of Neurog3 (a gene controlling pancreatic differentiation) in mouse...(trafficking to a subcellular location, start of gene expression, development of a cell structure, etc...formation of limbs are correlated with changes in gene expression. In a developing embryo, for example,...long-lived transcription factors control gross changes in gene expression as new body sections are formed. In synthetic...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...

...confounding mutations, the risk is high. You are generating a gene therapy product with CRISPR to (hopefully)...not to cleave DNA and instead activates/represses gene expression by various mechanisms. The RNA guide ...which involves making specific knock-out edits in a gene(s) of interest and has an average off-target frequency...involves making a large sgRNA library to thousands of genes and targeting and sequencing millions of cells. ...confound an experiment due to their location (e.g. in a gene in a similar pathway, a growth factor, etc.) this...Increasing the Specificity of the CRISPR-Cas System in Gene Editing. Mol Ther. 2021 Mar 3;29(3):937-948. doi... Gao F, Guo Y. Off-target effects in CRISPR/Cas9 gene editing. Front Bioeng Biotechnol. 2023 Mar 9;11:...

...LTRs, gag, WPRE, antibiotic resistance-genes, or the transgene itself. Many researchers prefer to design...multiplicity of infection (MOI) for predicting gene transfer events." Genetic vaccines and therapy 2.1 (2004): 1....with your virus and assaying for expression of a gene carried on the transfer plasmid or quantifying the...popular approach utilizes the antibiotic resistance genes carried on transfer plasmids. Target cells are transduced... to contain 1 integrated copy of the qPCR target gene. Developing the proper control line typically requires...Additional Resources on Addgene.org Learn How We Do Viral Production at Addgene Read Our Lentiviral Guide... that specific feature. Primers targeting the transgene are also beneficial as they ensure that the correct...

...Cpf1 multiplexing method, other multiplex CRISPR gene editing methods relied solely on Cas9. Overall, ...Lastly, an AAV vector was used to multiplex edit 3-4 genes in a primary culture of mouse neurons and in vivo...summary of multiplexing efficiency in vivo. Multiplex gene editing with CRISPR-Cpf1 is one of the latest developments...transfection, viral transduction) for simultaneous multi-gene editing with a single crRNA array. Are you interested...        References 1. Zetsche et al. "Multiplex Gene Editing by CRISPR–Cpf1 Asing a Single crRNA Array...review of Cpf1, check out this blog post or see Addgene's CRISPR guide page for a review of Cas9.  For a...plasmids; for multiple edits, single plasmids are generally more efficient. Pro-Tip Make sure to reverse the...

... certain guide sequences - using 5 guides/gene for 10 genes, they did not observe differences in cleavage... BELOW Biologists are going gaga over the newest gene-editing protein - a DNA-cleaving Argonaute from ... original sequence. If you’re looking to disrupt gene function via indel formation, this property could...Natronobacterium gregoryi, or NgAgo for short. Addgene has already distributed this plasmid all over the...supplied. Find Plasmids from the NgAgo Publication at Addgene Cas9 and NgAgo head to head NgAgo has a number ...27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan C., et al.  “DNA-guided DNA interference...PMC4697943. Find plasmids from this publication at Addgene. 3. Swarts, Daan C., et al. “Argonaute of the ...

...developed as well as CRISPR-based gene repression tools to find and test genes that may play a role in the ...proteomic tools to discover genes and proteins associated with pathogenesis. In his future work. Dr. Bengyella...in China Additional Resources on Addgene.org Learn About Addgene's Mission Check Out Our International...As we mentioned a few months ago, Addgene has been working with Seeding Labs to provide plasmid grants...and Duedu and how they plan to use plasmids from Addgene to advance their research. Louis Bengyella Dr. ...that enable E. coli and Citrobacter freundii to generate sugars from cellulosic biomass. Dr. Duedu would...would like to use the plasmids he acquires through Addgene to produce molecular biological tools like Taq ...

...CRISPRa components to adipose cells to activate genes involved in converting white adipose tissue (WAT... metabolizing glucose to dissipate heat. Of the genes they tested, activating UCP1 had the most potent...increasing oxygen consumption and upregulating other genes normally expressed in BAT. When donor-derived CRISPRa-AAV-treated... as well as extensive validation of the cell and gene therapy, so please check it out! Find AAV-CRISPRa...— partly because of its relative simplicity as a gene editing system. Despite that simplicity, its accessibility...method uses CRISPR interference (CRISPRi) to regulate gene expression of colorful, easily visible chromoproteins...CRISPR kit for equitable and accessible education in gene editing and synthetic biology. Nature Communications...

... Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible tagging:...tag. Selectable and stable: A puromycin resistance gene makes stable cell line creation a breeze. Modular...labeling By Alyssa Shepard To fully understand how genes are regulated, it's important to know what proteins...identify both new and known regulators at the FOS gene promoter and the MYC locus (Figure 5). Figure...identification of heat-shock proteins at the FOS gene promoter using TurboCas, streptavidin immunoprecipitation...very quick and efficient way to learn more about gene regulation. Find the TurboCas plasmid here! Cenik...Enhancer-AAVs targeting interneurons By Mike Lacy Targeting gene expression to specific cell types in the brain can...

...cell lines that stably express a gene of interest. While the general approach for using lentivirus, infect...multiplicity of infection (MOI) for predicting gene transfer events.” Genetic Vaccines and Therapy 2.1 (2004) Pubmed...Mycoplasma infection significantly alters microarray gene expression profiles.” Biotechniques 35.4 (2003)....Downstream processing of oncoretroviral and lentiviral gene therapy vectors.” Biotechnology Advances 24.3 (2006...production of lentiviral and retroviral vectors.” Human Gene Therapy Clinical Development 25.3 (2014). 175-85...lentiviral vectors through pseudotyping.” Current Gene Therapy 5.4 (2005). 387-98. Pubmed PMID: 16101513... cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting ...

...Precious Tools to Improve Gene Modification of Hematopoietic Cells for Research and Gene Therapy. Viruses 12...To deliver genes using lentiviral vectors, you need an envelope protein on the virus’s surface and a ... plasmid which contains a promoter, the envelope gene, and a polyA tail.   Figure 1: Lentiviral...can’t be used for neuronal tracing or peripheral gene delivery. Combining the rabies glycoprotein into...Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods 28:291–301 . https://doi.org/10.1089...Resources on Addgene.org: Check out the lentivirus guide Find ready-to-use lentiviral preps at Addgene ...your viral vector, you use the same protocol for generating lentivirus, but you use a different envelope ...

.... More than half of these sequences were generated by Addgene as part of our QC process. Automation and...From the beginning, one of Addgene’s goals was to ensure the scientific community obtained high-quality...the years to keep up with the latest tools. In Addgene’s early days, we would ‘spot-check’ critical regions...sequencing at our facility using Illumina MiSeq Next Generation Sequencing (NGS) technology. As our repository...will describe the importance of bioinformatics at Addgene and how our search for solutions led us to a partnership...bioinformatics needs       Fig. 1: QC'ing data at Addgene is like standing under a Niagara Falls sized rainbow...our software, even with continuous updates. The generous donation of a Illumina MiSeq sequencer enabled...

...intended. With the use of applications like SnapGene and Geneious, I was able to determine if any discrepancies...able to explore other parts of Addgene. This was done through the “Addgene 101” series, where each team ... Communication function at Addgene. Working a 9-5 As it turned out, Addgene uses something called “core...completing their co-op at Addgene. This is her second contribution to the Addgene blog.      More resources...brought me, a second year bioengineering student, to Addgene. I am currently working on the QC/QA team as a ...specific overview. It was immediately clear that Addgene wanted to ensure applicants understood what the...that the two hours were split up between several Addgene employees, who all made the daunting task feel ...

... at Addgene's booth, #3422, in the nonprofits section.  The Booth What will you find at Addgene's booth...scientific curation manager, Angela, experts on the Addgene repository, are also attending! Finally, our team....  You'll also find tons of information about Addgene's ready-to-use antibodies, viral vectors, and, of...course, plasmids! Plus, we'll be giving away fun Addgene swag for the duration of the conference (be warned...

...MTAs per year.  The MTA Process at Addgene As a repository, Addgene doesn’t actually own the majority ...distribution for their plasmids. MTAs at Addgene Since Addgene was founded in 2004, we’ve handled the majority...If you’ve deposited or requested plasmids from Addgene, then you’ve been involved in the material transfer.... The UBMTA, which is the MTA most plasmids at Addgene are shared under, is well-understood and accepted...for managing research use agreements like MTAs.  Addgene does a lot to ensure that we can facilitate a fast...signing of MTAs by the Provider and Recipient (in Addgene parlance, the depositor and requestor) organizations...that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer office is sent...