...proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed under native...sequence and a selectable gene product for isolation of successful transformants. Addgene distributes several...be interested in attaching epitope tags to their genes of interest, allowing for easy capture and detection...modifications in S. Pombe, including full and partial gene deletion, overexpression (by promoter substitution...few. Have you used these tools in your own lab? Addgene would love to hear from you, our community, about...

...526) that (1) enables efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity...for in vivo studies; (3) is compatible with single gene KO and pooled screening approaches; (4) encodes ... optimize conditions for a screen that assessed genes that regulate antigen-specific CD8+ T cell responses...screen, we used the pRDA_526 vector to knock out 30 genes putatively involved in CD8+ T cell responses to ...Markson et al., 2024), developed and deposited at Addgene by Arlene Sharpe’s lab, enables in vivo screens...10.1084/jem.20230699. Additional resources on the Addgene blog X-CHIME: Context-Dependent Germline Knockout...

...REVeRT, for gene delivery using reconstitution via mRNA trans-splicing. By splitting a large gene into smaller...the introduction of extraneous genetic material to reconstitute the gene of interest, reducing the immunogenicity...hIBA1a, 466 bp) that drives high microglia-specific gene expression when incorporated in an AAV vector (Figure...doi.org/10.1101/2023.12.12.571321.  Revolutionizing Gene Therapy: The REVeRT Dual AAV Vector System By Vaibhav...AAVs, and then splicing their mRNAs, the functional gene product is reconstituted within the target cells...vector-based therapies. REVeRT supports advanced gene expression regulation, enhancing both the mechanisms...trans-splicing dual AAV vectors for (epi)genome editing and gene therapy. Nat Commun., 14(1), 6578. https://doi.org...

...available at Addgene CRISPR-Cas9 References, Protocols, and more References: Generation of Gene-Modified ...the CRISPR/Cas9 system to precisely edit specific genes in monkeys. "Our study shows that the CRISPR/Cas9...system enables simultaneous disruption of two target genes in one step without producing off-target mutations...Gene-Modified Cynomolgus Monkey via Cas9/RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014...many human diseases are caused by genetic abnormalities, targeted genetic modification in monkeys is invaluable...invaluable for the generation of human disease models." Addgene Outreach Scientist and fellow blogger Caroline...

...variations in gene expression, they found that for approximately 40% of the identified genes, the genes were not...explanation. Next, they hypothesized that differences in gene expression could explain why some ancestry groups...groups were differentially dependent on certain genes. This didn’t prove to be correct, either. However, ...sequences for sgRNAs that target ancestry-associated genes.   What does the reference genome actually reference...cases, this may lead researchers to deprioritize genes that could be good targets for therapeutic development...cells’ germline genetic variations. They then wanted to identify cancer-associated genetic dependencies ...ancestry which will reduce the likelihood of generating genetic dependency data with an ancestry-based bias...

...on vectors where the gene of interest is inserted into the middle of the lacZ gene, thus disrupting 𝛽-...where only cells that have gained or lost a specific gene survive (ex: antibiotic resistance marker). In a... the blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which ...scientists have found that when a portion of the gene is deleted, it creates a non-functional 𝛽-galactosidase...mentioned above. In molecular biology, cloning a gene into your plasmid is often a numbers game. By using...troubleshooting Sanger sequencing results. At Addgene, we now use next-generation sequencing during our QC process....pigment. Find plasmids for blue-white screening at Addgene! Some bacterial strains contain lacZ in their genome...

...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...These vectors allow you to test the function of Your Gene Of Interest (YGOI) in a controlled environment under...design. You can create a new strain with a disrupted gene by transforming wild-type yeast with an integrating...modifications to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check... More Plasmid Resources from Addgene: Plasmids 101 blog series Addgene's Molecular Cloning Guide & Other... is which vector will best suit your needs? At Addgene, we have a vast collection of empty backbones that...vivo (try pcDNA3-EGFP). If you can get away with generating transiently transfected cells, you will not need...

...region of the gene they are trying to disrupt or delete, which both inactivates the gene and acts as a...maintain the plasmid. Adding an antibiotic resistance gene to the plasmid solves both problems at once – it...Historically, antibiotics have also been used to disrupt genes at the chromosomal level. Scientists introduce an...typically has two separate resistance cassettes and your gene of interest is cloned into/inactivates or completely...resistance!  What are antibiotics? Antibiotics are generally defined as agents that kill bacteria, or inhibit...lab, their mechanism for killing bacteria, and general working concentrations. For instructions on how...how to prepare antibiotic stocks, see Addgene's Reference Page. How else can antibiotics be used in the...

... gRNAs) that targets ~4,000 E. coli genes with ~15 gRNAs per gene and includes ~10,000 gRNAs targeting...CRISPR interference (CRIPSRi) platform for targeted gene repression in Candida albicans was developed by ... transcription of HSP90 an essential C. albicans gene. Read the paper in mSphere Find the dCas9 and... a few of the new CRISPR plasmids available at Addgene. We will still periodically focus on specific CRISPR...a new CRISPR tool you’ve recently deposited to Addgene and you’d like it to be included in the next What...more information. Additional resources on the Addgene blog Download the CRISPR 101 eBook Find our CRISPR...posts Read our CRISPR cheat sheet Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by...

...Energy’s Joint Genome Institute for uncharacterized genes that were proximal to a CRISPR array and the universal...CRISPR-Cas systems containing cas1, cas2, cas4, and a new gene cas14. cas14 encodes for a small Cas protein (40...-Cas systems. There are 24 variants of the cas14 gene that cluster into 3 subgroups (cas14a-c). All these...potential of Cas14, the Doudna lab cloned the cas14 gene together with the adjacent CRISPR array and intergenic...presence of infectious organisms (ex: SARS-CoV-2) and genetic mutations. By providing a gRNA specific of a viral...ID PMID: 30337455. Additional resources on the Addgene Blog Learn more about DETECTR Browse all of the...Download our CRISPR cheat sheet Resources on Addgene.org Find the Cas14 plasmids here Checkout our CRISPR...