...their test genes, the researchers at the BIOFAB showed that expression of many different genes was predictable...translational repressor to down-regulate a gene of interest. You can generate PUF proteins with a wide variety ...restriction enzymes and standardized primers to clone your gene of interest into both sequencing (pInitial) and ...abundance (meaning they are unlikely to be found in your gene of interest) and prevent incorporation of extensive...kit contain a variety of promoters to facilitate gene expression in bacteria, yeast, mammalian cells, ...hope this kit will allow you to quickly study your gene of interest in a variety of model systems, thereby...peptides) downstream of which you can clone your gene of interest. Translation of the standardized leader...

... one bacterium sharing genes with another, an accidental transmission of genes from one bacterium to another...‘horizontal gene transfer’. Resistance can also arise from random mutations in the genetic sequence that...Additional Resources on the Addgene Blog Learn How Antibiotic Resistance Genes Are Used in the Lab Probe...consuming to treat. Learn How Antibiotic Resistance Genes Are Used in the Lab How does antibiotic resistance...bacterial cells), or a bacterium picking up resistance genes from it’s environment (from DNA left behind after...develop, either through random mutation or horizontal gene transfer, means to defend themselves from these ...while the few that happen to possess resistance genes survive and multiply. Factors contributing to the...

... what part of a gene enhancers like to interact with, and how many promoters or genes each enhancer is...plot line in gene regulation. When chromatin conformation data is compared with changes in gene expression...dynamic role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated with CAPTURE...changes in gene expression and determine who’s a key driver of these changes. By generating a more vivid...updated Jul 2, 2020. Promoters may be the star of gene regulation, but enhancers and chromatin looping ... bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers are located ...located thousands of base pairs away from the gene they regulate, but are brought in proximity by the looping...

...viral (AAV) vectors are the most frequently used gene-transfer tools in the study of the brain and spinal...organs to the CNS, are also an important target for gene delivery, especially for the study of pain. While...all possible combinations into a region of the cap gene. In this special CREATE cap plasmid, loxP sites ...primers to determine which version(s) of the capsid gene successfully infected the cell type of interest....translational applications (mechanistic studies and gene therapy treatments for neurological diseases). The...capsids can serve as powerful cell-type-restricted gene vectors for neurons, no transgenic mouse required...et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems...

...resistance gene, allowing resistant bacteria to multiply, spreading their resistance genes throughout ...them to multiply and propagate their resistance genes. Our society is hoping to combat antibiotic resistance...bacterial species, or even an antibiotic resistance gene. Importantly, native, non-pathogenic bacteria remain.... targeted beta-lactam and quinolone resistance genes in E. coli using plasmid and phagemid delivery systems...population to the antibiotic, despite this resistance gene being encoded by a high-copy plasmid. In the case...CRISPR machinery targeting 1) the given resistance gene and 2) a second lytic phage. Bacteria that receive...work targeted two distinct beta-lactam resistance genes, again with very high specificity. This double phage...

... containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral ...Talking 'bout my (lentiviral) generation The first step in delivering genes via lentivirus is deciding what... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges...split the viral genome by removing the viral Rev gene from the packaging plasmid and incorporating it ... the structural elements, envelope, and transfer gene are transfected into a packaging cell line, usually...) while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how...sufficient titer; this is often the case with large genes. This is due to inefficient packaging of large inserts...

...screens using CRISPR technology, gRNAs to knockout genes – are essential screening components, the model ...Sometimes, you might have a few positive control genes on which to test the validity of your biological... a new assay or both, and you have no idea what genes are involved. Or you don’t know how well your assay...definition, the kinome is a well-annotated collection of genes that can provide a useful starting point for developing...screens can be performed with either 4 or 8 gRNAs per gene. Further, the pools are available in two vector ...individual plasmids will also be useful for imaging or gene expression assays necessitating the screening of...blogger, member of the Addgene Advisory Board, and Associate Director of the Genetic Perturbation Platform...

...was previously a large barrier for gene therapy. In fact, AAV gene therapies have already been approved...Transient Gene Expression High Immunogenecity AAV Remains Episomal (Little Risk of Mutagenesis) Very...of Insertional Mutagenesis) High Gene Expression Infects Non-Dividing Cells Large Packaging Capacity (...Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy with...Type of Virus Pros Cons Retrovirus Stable Gene Expression Broad Tropism Only Infects Dividing...Immunogenicity Infects Non-Dividing Cells Transient Gene Expression Low Tropism for Mouse Cells and Hematopoietic... lentiviral vectors through pseudotyping.” Curr. Gene. Ther. 5 (2005) 387-398. PubMed PMID: 16101513. ...

...of homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids used...host mutations by including a copy of a functional gene which complements the host’s auxotrophy. When grown...have incorporated the plasmid carrying the required gene.  The table below lists some of the most commonly...growth-based positive selection for the loss of the marker gene) For use in: Can this be used in other yeast species...yeast phenotypes. This marker should not be used in gene disruption experiments. MET15 L-methionine and...FOA in fission yeast induces mutation in the ura5+ gene in addition to ura4+. leu1+ L-leucine   S. pombe...issues. One method to reduce the amount of marker gene expression is to use a partially defective promoter...

...explains how gene drives work and what work he's planning on doing with CRISPR-based gene drives focusing...developing new CRISPR tools, and the potential uses of gene drives. Listen here! Podcast breakdown (Time : ...- 31:25 : Nature's defenses against gene drives. 31:26 - 33:08 : Plans for future work at the Wyss Institute... gene drives...became a member of the Addgene advisory board where he will help shape Addgene's future. Listen to this...member of the Addgene Scientific Advisory Board with a particular interest in generating Cas9 based tools...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     Additional...