What? You want to do this the hard way?
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!
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