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An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

Posted by Tyler Ford on Dec 8, 2015 10:30:00 AM

We recently sat down with Addgene Board Member, depositor, and Harvard Medical School researcher Connie Cepko. Listen to the podcast below to hear all about the research being conducted in Professor Cepko's lab and to get some insight into her management and mentoring styles. If you'd like to learn more about Professor Cepko's recent work on developing GFP-activated proteins, read our recent blog post.

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Topics: Interview, Viral Vectors, Podcast

New Tool for Lineage Tracing: The ClonTracer Library

Posted by Tyler Ford on Sep 22, 2015 10:30:00 AM

This article is based on an interview with Novartis researcher, Carrie Bhang.

The ClonTracer Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis Oncology, is an exciting new tool that allows researchers to individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and clonal dynamics over time using next generation sequencing (NGS). The library was developed when Carrie was a post-doc in Frank Stegmeier’s lab in Novartis Oncology. 

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Topics: Interview, Viral Vectors, pooled libraries, Cancer

A Match Made in Heaven: CRISPR and AAV

Posted by Mary Gearing on Jul 14, 2015 10:30:00 AM

This post was updated on Dec 4, 2017.

CRISPR genome editing has quickly become the most popular system for in vitro and germline genome editing, but in vivo gene editing approaches have been limited by problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery due to their low immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, packaging Streptococcus pyogenes (SpCas9) and a chimeric sgRNA together (~4.2 kb) into an AAV vector is challenging due to the low packaging capacity of AAV (~4.5 kb.) While this approach has been proven feasible, it leaves little room for additional regulatory elements. Feng Zhang's group previously packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating purification and co-infection of two AAVs.

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Topics: CRISPR, Viral Vectors

Plasmids 101: Viral Vector Elements

Posted by Marcy Patrick on Jul 17, 2014 3:09:00 PM

The use of viral vectors in research is beneficial for a number of reasons, including but not limited to: helping to get difficult-to-deliver DNA into mammalian cells, increasing the efficiency of gene transduction, allowing for control over which cells are infected through viral pseudotyping, and ease of vector cloning and modification. At the most basic level, viral vectors consist of a viral genome that has been adapted into a plasmid-based technology and modified for safety through the removal of many essential genes and the separation of the viral components. Read on for a brief description of the viruses used to make these vectors as well as a table defining the major elements found within the plasmids comprising the viral vector systems.

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Topics: Plasmid Elements, Plasmids 101, Viral Vectors

Your Lentiviral Plasmid FAQs Answered

Posted by Kendall Morgan on Apr 23, 2014 9:08:00 AM

Lentiviruses are useful and efficient tools to introduce your gene of interest into cells. Unlike gamma-retroviruses that can only infect dividing cells, lentiviruses can infect dividing and non-dividing cells. 

Addgene has an extensive collection of lentiviral plasmids created for a variety of applications including cDNA expression, shRNA-mediated knockdown, Tet and Cre-regulated expression, CRISPR genome editing, and more. Not surprisingly, we receive many questions from scientists all over the world looking for some additional information or clarification on these vectors. Read on to find the answers to our most frequently asked lentiviral questions.

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Topics: Plasmid How To, Plasmid Elements, Lab Tips, Viral Vectors

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