...interested in heftier genes, you can coinfect with multiple viruses to express genes or gene sets over 4.7 kb...capacity of ~ 4.7 kb, meaning that your gene of interest (or genes) need to be 4.7 kb or less to fit in ...expressing multiple small genes, you can design single rAAV vectors with multiple genes packaged in them and...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression ... transient gene expression in a particular cell type to CRISPR genome editing, optogenetics, and chemogenetics... AAV is delivery of genetic cargo to a host cell. In order to make room for a gene of interest, AAVs used...icosahedral capsid, that can be used to express genes of interest in cell and animal models. AAVs are ...

...isoform-specific gene expression. Gene isoforms refer to mRNAs generated from the same gene but with different...isoform-specific gene regulation — at least, more straightforward than generating cDNA for each gene isoform individually...first ask yourself whether gene overexpression is necessary (instead of gene knockout or knockdown, for...tools for modulating gene expression in the central nervous system (CNS). To address this technological ...combinations of transcriptional activators to induce gene transcription at genomic sites targeted by specific...libraries. However, using CRISPRa to target individual genes can be relatively laborious because multiple plasmids...packaged into AAV for in vivo delivery. Multiplex gene targeting is also possible and is only constrained...

...time-consuming, as it only changes one gene at a time. CRISPR-based systems enable gene knockout in immune cells in... to create mice with gene knockouts in their immune systems. However, many genes have different functions...combination with different genes. Having CRISPR systems for assessing genes in defined contexts would ... deposited at Addgene by Arlene Sharpe’s lab, allow for wide deployment of in vivo gene knockout in immune...Annotation of genes in immune cells typically involves the creation of germline knockout mice, which ...the constitutive, ubiquitous knockout of single genes in immune cells in vivo (LaFleur et al., 2019a and...Mouse Strain C-CHIME Knocks out a pair of genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9...

...two genes: the replication (rep) gene and the capsid (cap) gene. In wild-type AAV, these two genes are...with BioRender.com   Replication (rep) gene The rep gene encodes four proteins that are required for... is called cross-packaging.  Capsid (cap) gene The cap gene encodes three AAV viral capsid proteins VP1...which contains the AAV structural and packaging genes, the adenoviral helper plasmid which contains the..., freeing up space in the viral genome so that a gene of interest can be packaged into the virus instead...Next let’s take a closer look at the rep and cap genes. Figure 1. The RepCap plasmid and its products...and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene expression is often driven by the AAV p5 or p19 ...

...both introduce genes into an organism’s genome (see figure) and to disrupt endogenous genes at the site ...first-generation transposase (as determined by transposition assays of antibiotic-resistance genes conducted...). In their Nature Genetics paper, the lab showed that efficiencies of stable gene transfer using SB100x... the rapid generation of stable transgenic cell lines which can robustly express your gene-of-interest... exciting tool for producing specific genetic mutations and gene disruptions without the use of viral ...transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-61. Epub 2009... et al. A new approach to gene therapy using Sleeping Beauty to genetically modify clinical-grade T cells...

...planarian genes, we encourage you to deposit them back with Addgene and expand the number of genes that other...expression of specific gene expression. The 49 deposited plasmids enable students to study gene expression and...planarians. For example, the gene Piwi labels neoblast cells and the gene Prohormone convertase 2 (pc2...beta-catenin. Other gene targets do not require amputation to be visible. For example, odf2 gene knockdown results...and functions of key regeneration-related genes in four inexpensive and commercially available planarians...types and for viewing how gene expression changes through time or after genetic manipulation. The technique...Scientists routinely use techniques to alter gene expression or to label specific cells, but there are...

...antibiotic resistance genes. Genes that are targets of antibiotics are often essential genes which creates a.... ICEBs1 carries the genes needed for conjugation, but can also carry cargo genes, like Mobile-CRISPRi...if that gene is a target of an antibiotic. For example, let’s take a look at the essential gene folA which...targeted essential and non-essential genes with two guides per gene. The library was grown in glucose minimal...essential genes were less represented while bacteria containing guides targeting non-essential genes remained...does not disrupt gene function and is stable and active for greater than 50 generations. Figure ...tools scientists can use to study them. For example, gene knockdown with CRISPR interference (CRISPRi) has...

...replication and viral gene transcription (6). The EBOV genome contains seven viral genes, each flanked by ...sequence (gene start) and a transcription termination/polyadenylation signal sequence (gene end). A minigenome...Figure 1). All viral genes are removed and replaced by a single (non-viral) reporter gene (e.g. firefly luciferase...luciferase or eGFP) flanked by the EBOV-specific gene start and gene end sequences. Thus, the minigenome contains...Cloning the viral genes in this plasmid would be more difficult compared to if each gene were encoded in...transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the gene start signal and ...of the reporter gene. The polymerase will elongate the mRNA until it reaches the gene end signal, where...

...depending on its orientation to your gene. T7lac High levels of gene expression Promoter from T7 bacteriophage...modulating gene expression through varied inducer concentrations. trp High levels of gene expression...human or mouse) General expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive...deacetylation. Ubc General expression mRNA Mammalian promoter from the human ubiquitin C gene Constitutive  ... human beta actin General expression mRNA Mammalian promoter from beta actin gene Constitutive  Ubiquitous... Requires the presence of Gal4 gene to activate promoter. Ac5 General expression mRNA Strong insect ...Actin 5c gene Constitutive  Commonly used in expression systems for Drosophila. Polyhedrin General expression...

...the env gene containing the rev response element (RRE), an internal promoter, and the desired gene (transfer...-1 genome containing all viral genes with the exception of the env gene (packaging plasmid), and 3) the...used to express your gene of interest either constitutively or inducibly. If your gene of interest is toxic...expression of your gene. The addition of Dox in the culture will then repress your gene of interest. This... Youichi Suzuki (2011). Gene Regulatable Lentiviral Vector System, Viral Gene Therapy, Dr. Ke Xu (Ed.)...harmful genes into a transfer vector. Lentiviral particles expressing proto-oncogenes or oncogenes for instance...safe use in a clinical setting. In this generation, the HIV Tat gene (previously used to drive expression...