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Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

Posted by Jennifer Tsang on Sep 4, 2018 8:53:28 AM

Researchers express genes of interest from plasmids in order to study gene function or to engineer cells for specific purposes. Unfortunately, plasmid copy numbers vary within cell populations and over time resulting in variable gene expression that can impact observed phenotypes. Factors such as the growth medium, growth temperature, and growth rate can all impact plasmid copy number in a cell.

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Topics: Plasmid Technology, Synthetic Biology, Plasmid Elements

Finding nucleic acids with SHERLOCK and DETECTR

Posted by Alyssa Cecchetelli on Aug 30, 2018 8:28:06 AM

Sensitive and specific nucleic acid detection is crucial for clinical diagnostics, genotyping, and biotechnological advancements. Current methods of nucleic acid detection however, either lack the sensitivity or the specificity to detect nucleic acids at low concentrations and/or are too expensive, time-consuming, and complex to use outside of standard laboratories. Recently scientists have utilized CRISPR-Cas9 protein variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect nucleic acids at the atomolar level.

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Topics: CRISPR, Genome Engineering, Plasmid Technology

Split-BioID: An Improved Method for Studying Protein-Protein Interactions

Posted by Beth Kenkel on May 1, 2018 9:57:26 AM

One way to define a protein’s purpose is by its protein-protein interactions (PPIs). These interactions are often modeled as binary relationships, i.e. protein A interacts with protein B; but proteins are social biomolecules. They can be part of multiple dynamic and overlapping complexes that have distinct functions. Many existing methods for identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability to specifically identify proteins that interact with a particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent biotinylation technique BioID. The key advantage of Split-BioID is that it allows for the validation of a binary PPI as well as the identification of additional interacting factors.

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Topics: Plasmid Technology, Hot Plasmids

RANbodies: Reporter Nanobody Fusions

Posted by Beth Kenkel on Apr 10, 2018 8:56:44 AM

Antibodies are a go-to tool for detecting a protein of interest in cells and tissues. Although antibody production is well established, it’s also a process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the Sanes Lab overcomes this limitation and allows for the flexible design and small scale production of antibodies.

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Topics: Plasmid Technology

CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

Posted by Guest Blogger on Feb 13, 2018 9:51:55 AM

This post was contributed by guest blogger Matthew J. Niederhuber, a graduate student at UNC Chapel Hill.

Chromatin immunoprecipitation followed by high-throughput sequencing, ChIP-Seq, is the go-to method for mapping where a protein binds genome-wide, and has been widely applied in many model organisms and cell lines. Although ChIP-seq is a relatively simple and robust protocol it does have limitations. The enzyme-based CUT&RUN method overcomes many of these limitations and makes it easier for you to map protein-DNA interaction with limited biological materials.

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Topics: Plasmid Technology, Techniques

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