Sigma BioBlogs

We are proud to let you know that you can now get a CompoZr Knockout Zinc Finger Nuclease to permanently knockout any gene in the human genome.

We are so knockout happy, that we found fighting genes.  Knockout APOLLO just like ROCK(y)1.

Or you could go for some old classic genes like  P53,  or Interleukin 1.

ANY gene in the human genome is part of our line of over 20,000 CompoZr Knockout ZFNs.

Each is functionally validated, ready for immediate transfection, and you can generate a knockout cell line in as little as 4 weeks.

CompoZr Knockouts are here! Learn more!

To celebrate…we have a short video to show you that not all knockouts are bad…

Learn more here from Dr. Dana Carroll of the University of Utah

For a contest earlier this year, we posed the question “Where did your Bio Begin?” and you answered with a creative collection of photos, stories and videos to demonstrate your passion for biology.

Dr. Sleutel’s lab was one of the grand prize winners who received a Custom Zinc Finger Nuclease.

The Sleutel Lab in their Bio-shirts!

This creative group won for their entry Science is addictive like M(aterials) & M(ethods).

We caught up with the Sleutel lab to learn more about their research, the inspiration for the video and what they plan to do with a Custom Zinc Finger Nuclease.

What is the focus of your research?

We are focusing on the fundamental question of how our genes are regulated and how this regulation can be influenced in the context of the packaged chromatin in the nucleus. Specifically we have been studying the CTCF protein family. These proteins contain 11 zinc fingers that in different combinations are used to bind to DNA. Binding to DNA generates DNA loops such that depending on the loop context genes can be switched off or on. We investigate CTCFs chromatin looping, effects on gene regulation and other epigenetic results using Embryonic Stem cells and mouse models in which we have disrupted and tagged genes and loci by gene targeting (using conventional homologous recombination strategies)

What do you plan to do with the ZFN?  How will it benefit your work going forward?

Still here we are brainstorming. But this technology allows us to manipulate genes in a cellular context rather flexibly.  More to come!  Sigma Bio Blogs will follow-up once the lab has more information on their use of the ZFN.

What was the inspiration for your video?

We have a Ph.D. in the lab, named Suzie, who is very fond (not to mention the word addicted) of the M&M chocolates. Her thesis deals with the function of the 11 zinc finger protein CTCF. Having our technician Michael dress up as an M&M was the hilarious start of the story in which he plays Suzie who has ups and downs in her Ph.D. project.

How did you develop your video submission?  Who were the brains behind it?

The video was entirely shot on the Erasmus Medical Center and mostly in the lab. The brain behind the video was Group Leader Frank Sleutels. Every year, he inspires and pushes the lab members to join in making an absurd, crazy and inspiring silly science video. There is no better way in team building than making such a video. Some of these can be found on Youtube.  A couple of these videos were shown on Dutch TV and he even got contacted through Youtube by the Discovery Channel.

To stay on top of technology we are planning to make this year’s video in 3D.  So stay tuned!

For now, you can view their winning video.

Welcome to the 3^rd blog in our series about Dr. Dana Carroll of the University of Utah.

The previous blogs focused on Dr. Carroll’s thoughts on Sigma’s role in ZFNs for genome editing, and on Scientists he admires.

In this video, we asked Dr. Carroll to share his thoughts on the current state of zinc finger nucleases, and where the technology is headed.   He explains how ZFN’s are being used in a broad range of organisms, from Zebrafish to mosquitos.  Then he comments on current clinical trials and the potential for ZFNs in plants.

Welcome to the second blog in our series about Dr. Dana Carroll at the University of Utah!  Dr. Carroll helped develop the Zinc Finger Nuclease (ZFN) technology for targeted genome editing offered by Sigma Life Science as CompoZr® ZFN.

In the following video, we asked Dr. Carroll to expand upon his thoughts about Sigma Life Science’s role in enabling genome editing through the use of ZFN technology.  He kindly praises the scientists of Sigma who have worked closely with Sangamo Biosciences to develop the best ZFNs available, and for taking this technology further by developing genetically modified cell lines and rodent models.

He also challenged us to continue making the nucleases more accessible to the larger scientific community. We have been working hard to meet his challenge. Access our CompoZr ZFNs, without breaking the bank.

Since we are a bit ZFN crazy right now with the airing of our ZFN Webinars, we wanted to feature the newest product based on the zinc finger nuclease technology, the CompoZr® Targeted Integration Kit (AAVS1).

Of course, you may have reasons why this kit isn’t for you so we think it is time for a top 5 list, our first on SigmaBioBlogs.

Reasons NOT to use the CompoZr Targeted Integration Kit:
1. You aren’t in a hurry to publish your results.
We are talking about the possibility of getting rapid biallelic insertion through a single transfection in as little as 48 hours….

2. You don’t mind inefficiency.
Genome editing using homologous recombination has served the research community well for the past two decades but it is highly inefficient.  A one in a million event, this method also requires sophisticated selection strategies.  The well-validated ZFNs within the CompoZr Targeted Integration Kit cut the DNA with high efficiency at the AAVS1 site.  The success rate is up to 3 to 4 orders of magnitude higher than spontaneous homologous recombination events.

3. You want varied expression levels and stability.
Previous methods to create transgenic cell lines use random integration of a plasmid.  The result is greatly varied expression levels and expression stability.  Use of a single genomic locus means stable, uniform expression levels.  We chose the AAVS1 site, a safe harbor locus that is ubiquitously expressed and its disruption leads to no adverse effects on the cell.

4. You like complexity.
We all know people like this and you may be one of them.  You can’t help but give a dissertation when a simple Yes/No answer would have sufficed.  If this is you, then this kit is just too simple for your taste.  Gene integration is at a single endogenous locus. The need to engineer cell lines with transgene landing pads is eliminated.  I mean, come on, that was the fun part, right?

5. You want us to design a custom ZFN!
We didn’t believe your other excuses but this is a reason we can rally behind.  We can rapidly design, assemble and validate a ZFN pair targeting YOUR gene of interest.

Now that we’ve established that reasons 1-4 don’t apply in your case, let’s go back to #1.  You DO want to be the first to discovery (this is what we love about you).  Did you know that the recently released Targeted Integration Kit could have been in your hands a year ago?  Participants in our Validation Testing Program obtain early access to some of our coolest products!

Read what one tester had to say about the program:
I was impressed by the validation testing program for a number of reasons. I had frequent and prolonged phone and email conversations with multiple scientists at Sigma and they listened intently to my input. Further, they went back and redesigned the kit a few times, trying to address where we (and others) had problems. It was clear to me that Sigma very stringently tested this kit to make sure that it will work as advertised.

Researchers who tested the CompoZr Targeted Integration Kit conducted our desired protocol using just 1/5 of the kit!  That is a lot of remaining product for your research.  Also, rapid responders receive incentives (such as gift cards) in addition to free product.  Find out more!

What would you say is as exciting as offering a novel game-changing technology?  Learning about that technology from the talented individual who helped to shape it!  Sigma Life Science is fortunate enough to have Dr. Trevor Collingwood as a member of the team.  Formerly of Sangamo Biosciences, Dr. Collingwood led the Enabling Technologies Group, focusing on development of the zinc finger nuclease platform for genome engineering.

If you are asking the question “What are zinc finger nucleases?” then you are in luck!   Sigma Life Science has created a series of FREE, live online seminars to introduce you to the technology and to delve into specific applications of zinc finger nucleases.

If you missed the introductory webinar held on May 6, the recorded webinar Introduction to Zinc Finger Nuclease Technology will be available for you to experience at your convenience.  We spent 30 minutes listening and this is just a taste of what we learned.

1.  What are ZFNs?
ZFNs are engineered DNA-binding proteins that act as “genomic scissors” to create a highly targeted double strand break in genomic DNA at any specified locus. The double strand break is repaired by the cell, leading to gene knockout or gene modification only at the intended locus. This technology will knock your genes right off!

2.  Design consideration
A two pronged approach of bioinformatics and protein engineering maximizes the specificity of ZFNs. A sophisticated algorithm is used to look for unique sites within the genome (target sequences that do not have homologous sequences even with 1-4 base pair mismatches) and avoid regions such as repeat elements, SNPs and splice variations. The Fok I nuclease is also engineered to function only when two different zinc finger nucleases come together, increasing specificity. Nothing like a clean break!

3.  Using ZFNs
Dr. Collingwood will cover the various applications of Zinc finger nucleases for targeted gene knockout and target integration.  The overall advantages for using zinc finger nuclease technology in both approaches include: modification of endogenous loci, ZFNs expressed transiently, no selection required (it is that specific!) and monoallelic or multi-allelic disruption/insertion.

Are you ready for more advanced topics in Zinc Finger Nuclease Applications? Of course you are.

May 27, 2010 - Gene Knockout in Cell Lines
Dr. Collingwood provides examples of both single and multiple gene knockouts drawing from work with transformed cell lines, primary human cells and multi-potent stem cells.
9 AM Central Time
4 PM Central Time

June 15, 2010 – Targeted Gene Integration
Join Dr. Collingwood for a discussion of various applications of gene integration including gene tagging (such as infusing GFP directly to the endogenous gene) and modification at the endogenous locus to recreate or correct disease-related mutations.
9 AM Central Time
4 PM Central Time

Register to attend the live webinars or check back at sigma.com/zfnwebinar for the recording to be posted.  We hope to have the recording of the first webinar posted soon.  Until then, check out this video on ZFNs.

CompoZr ZFN Technology for Targeted Genome Editing

It’s with great pride that we tell you Sigma Life Science is Where bio Begins.

To quote our VP of Marketing, Dr. Helge Bastian…“We want to help life science researchers address their research challenges and are committed, more than ever to support them on their way to new and revolutionary discoveries.”

Over the past decade, Sigma Life Science has built a repertoire of products and technologies to support the biological researcher. Part of our commitment to biology is to share our knowledge and resources with you.

If you’ve been following us over the last 30+ days, you have noticed that we are using social media as a means to get to know the biological community at a whole new level. We’ll continue this trend here. As we go forward look to sigmabioblogs.com for the skinny on new techniques, improving old techniques, exciting applications, war stories from technical services, and information about scientists from all over the bio community.

Here is a video of our President, Dave Smoller, to tell you the details.

Watch the video and then check out our great tools and techniques listed below.

• Targeted Genome editing
• Knock out research models
• Custom oligo services
• Biomolecules including antibodies and small molecules
• Reprogramming tools for stem cell research
• siRNA and shRNA’s for gene knockdown
• Biological search tool

Thanks for talking to us. We are having fun and hope you are too.

Welcome to Sigma Life Science. This is Where Bio Begins.