...an unpublished set of Zika gene plasmids to Addgene’s repository. The genes, from the Zika virus Asian...vectors targeting their genes of interest to generate mAID fusions to endogenous genes. Cell lines with the...targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences can...expression level for their gene of interest. Although the parts involved in altering gene expression are relatively...contains ~90,000 gRNAs that target ~15,000 genes with 6 gRNAs per gene. The second is a "supplementary" library...library to generate a population of HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA... greatly simplified the process of generating multiple E. coli gene expression plasmids with the Cross-disciplinary...

...been used for many years for in vivo delivery and gene therapy and represent one of the most studied viruses...CRISPR-Cas. Figure 1: Schematic of targeted gene knock-out (left) and targeted DNA knock-in (right...binds to target DNA. In the final step of targeted gene knock-out (left), a fragment is deleted from two... This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing the delivery... CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various cell types...

...efforts to regulate gene expression were limited to inducible promoters or direct gene knockout. In contrast... of gene expression. They successfully knock down expression of essential and non essential genes in a.... These could be genes, specific polymorphisms, including antibiotic resistance genes. This approach successfully...resistance pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance E. coli carrying...wild-type sequence persists, Cas9 will cleave the gene of interest, inducing a lethal DSB.     This system.... pCas contains Cas9 and the phage recombination gene lambda Red. pTargetF contains the specific gRNA(... cleavage induces HDR to directly modify a given gene. CRISPR-tranposons By combining CRISPR editing and...

...since become a gold standard in gene expression analysis and a luciferase gene (one of many available to choose... Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves as.... The effect of promoters or enhancer regions on gene expression can be determined by detection of the...effect of mutations of these regulatory elements on gene expression. The regulatory element (wild-type or...or mutant) is cloned upstream of the luciferase gene in the plasmid, which is then introduced into the... cells. Expression of luciferase as the reporter gene is measured via its activity to produce light, often...measurement of miRNA efficiency on a target sequence. The gene of interest is cloned into a plasmid downstream ...

...this plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY...already have a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform ...co-transform pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled... Resources on Addgene.org Check out the Molecular Biology Reference Download Addgene's Plasmids 101 eBook...simply more precise. “We show that you have less heterogeneity in the population and you can do all of these...permission.   BLADE is just one of the several optogenetic tools that Di Ventura’s lab and Khammash's lab...lab have created and shared with Addgene. You can find all the plasmids deposited from Di Ventura's lab...

...Adeno-Associated Viral Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https...Specificity of gfaABC1D-Driven Transgene Expression in AAV-Mediated Gene Transfer. Molecular Therapy - ...easier to do the validation first.  AAV-mediated gene expression can differ extensively depending on the... cells in far away brain regions expressing your gene of interest. This can cause issues in tracing studies...can occur. Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous...chicken-beta-actin or CAG can drive strong early gene expression, but can become methylated over time,...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig. 3) ...

...cost-effective manipulation of gene expression in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https...system, in which the required adenoviral helper genes are supplied on two different packaging plasmids...pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing replication ...replication factors and serotype-specific capsid genes.  In this helper-free system, HEK293T cells, expressing Ad...transfected with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly...derived inverted terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org...genomes can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038...

...using AAV as a tool for gene expression in mammals. Resources on Addgene.org Search for AAV vectors ...One of the biggest barriers to gene therapy treatment of central nervous system (CNS) diseases is the...has not been identified. While LY6A or other Ly6 gene family members are linked to viral susceptibility...large phenotypic effects such as those that affected gene expression, mRNA splicing, or protein coding regions...as non-permissive to PHP.B CNS transduction. Two genes were associated with the PHP.B permissive phenotype...permissiveness. The Wilson Lab also looked for the gene variant responsible for the PHP.B permissive phenotype...inheritance pattern that suggested there was a single gene that had two codominant versions or alleles that...

...for doing gene targeting, reporter generation, controllable gene knockouts, and inducible gene inactivation...Alec Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, developed by Yiping... Shirley Liu's lab targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize... to develop CRISPR-based approaches for studying gene function in these systems. As a part of this effort...Yiping Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...opportunities for investigating genetic mechanisms related to morphogenesis, cell fate specification, and...Lab have compiled a new toolkit for performing genetics studies in organoids. The Organoid EasyTag System...

...the Grousbeck Family Chair in Gene Therapy, Director of the Grousbeck Gene Therapy Center, Associate Director...nonprofit companies. His lab focuses on gene therapy work for genetic diseases and over the past few months...deposited AAV constructs for gene delivery. In 2018, he interviewed Addgene co-founder Melina Fan and former...PhD Khandan Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development,...Dimension Therapeutics, she helped build and grow the gene therapy division of the company.  Ultragenyx’s goal... systematically test the function of all 20,000 genes in disease models and then focus drug discovery ...to crises like those and ended up in the field of gene therapy.” He’s also worked on vaccines throughout...