...the sgRNAiBAR library covers every annotated human gene, it enables genome-wide screening with a high MOI...a stop cassette upstream of an effector/reporter gene. In cells where the stop cassette is removed, the...fish, and mice, by replacing the effector with a gene of interest in the cells you are studying. Read ...calcineurin-dependent response elements (CDREs) and drives gene expression. Because calcineurin is required for ...antifungals that target calcineurin. Current calcineurin gene reporters are based on the bacterial βgal assay,...Competition (SPARC) is a fully genetic tool for labeling just a fraction of a genetically-defined subset of cells...lentiviral vectors the iBARs can be detected by next generation sequencing.  In order to avoid a high false-positive...

...sensitizing gene cassettes. Thse vectors are engineered to replace the drug resistantance genes in the bacteria...were identified for 61 such diseases. Now single gene alleles have been identified for 4,163, over 50%...Lowe talked about his work on the 500+ human tRNA genes and, until now, we've overlooked regulatory fragments...may be involved in preventing silencing of self-genes. Dr. Mello presented a hypothesis that these pathways...involved in RNA based surveillance of repetitive gene families and provide a way to guide inheritance ... field. Resources at Addgene Browse Addgene's RNAi resources Browse Addgene's CRISPR resources Read ...countries in number of plasmid requests from Addgene. Addgene - Canton Nucleic Acids Forum 2015 from Joanne...

...Ehrhardt, A., et al., Episomal vectors for gene therapy. Curr Gene Ther, 2008. 8(3): p. 147-61. PubMed PMID... functional genes at the site of insertion and which can potentially promote tumorigenesis in vivo. The...fellow at Massachusetts General Hospital with research interests in the fields of gene editing and stem cells...non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures. Cell Res, 2011. 21...The phenomenon has been recorded in past gene therapy trials and remains a safety concern in the clinical...clonal T cell proliferation in two patients after gene therapy for SCID-X1. Science, 2003. 302(5644): p...the chances of insertional mutagenesis, which is the integration of transgenes into the genome that can ...

...used for genome editing, activating gene expression, repressing gene expression, and much more. But there...engineering viruses. For example, an Acr gene was used to replace genes in the difficult-to-engineer Sulfolobus... Selection marker while engineering viruses Acr genes can be used as positive selectable markers in engineering...et al., 2018). Only viral particles that have the gene deletion could replicate when challenged with the...Experiment Read more about Optogenetic Control of CRISPR   Resources on Addgene.org Find CRISPR Plasmids ...Nme) (Pawluk et al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and well studied...10.1126/sciadv.1701620 Additional Resources on the Addgene Blog First time CRISPR user? Check out this post...

...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...

...within the gene/insert, this information can be found here. There may be more than one gene inserted into...can easily clone my gene into it? Your best bet is the Plasmid Map: We auto-generate a plasmid map based... deposited with Addgene, you can list the original vector’s plasmid number here. Gene/insert: This describes...the backbone, and provides information about this gene or DNA element of interest. If the depositor has... into a backbone, and each is given its own gene/insert section on the plasmid page. Tags are listed according...We Do Quality Control at Addgene Resources on Addgene.org Join the Addgene Community Learn the Benefits...plasmid. Use the Addgene catalog number to easily find template citations for Addgene plasmids you have...

...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. ...

...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...

... 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)....

...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...