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Frequently Asked Questions (FAQs)
What is SilenciX cell line?
The SilenciX cell lines are a ready to use system for studying
specific silencing of a gene of interest. For each reference a gene has
been silenced. For example, ERCC1 HeLa SilenciX are cells in which the
endonuclease ERCC1 involved in DNA repair has been extinguished.
My target is not listed in your catalog, how to benefit from
SilenciX technology?
If you would like to work in HeLa cells, we will prepare SilenciX as a
catalog product. You would agree that this cell line will be put on
the catalog to benefit from our list price.
What is the delivery time of a SilenciX preparation?
70-90 days maximum to silence a target gene in Human Hela cells.
Can I buy the vector separately?
No, it is not possible. Only SilenciX cell lines are available.
I am working with Mouse cell lines, can you perform SilenciX
cell line for me?
The SilenciX approach is available only to Human cells, as the benefits of the silencing vector are based on human promoters.
Do I need a particular safety handling for the use of SilenciX?
Although shRNA expressing vectors contain small elements of Epstein-Barr virus, they do not have the capability to produce viral particles (unlike adenovirus- or lentivirus-based plasmids). However, handle as a potentially bio-hazardous material under at least Bio-safety Level 2 (BSL-2) containment conditions. You are responsible for its safe storage, handle, and use. SABiosciences Corporation is not liable for damages or injuries resulting for its receipt and/or use.
TARGET
How do you design shRNAs?
The design for shRNA is based on an algorithm predicting effective
siRNA with specific rules developed for optimization.
Can I have access to the shRNA sequences?
The shRNA sequences can be sent to you on request.
Can I have my own specific target gene silenced? in my own cell
line?
shRNA sequence can be designed for any Human target gene. We provide
HeLa SilenciX cell lines as a catalog product or any Human cell line
you want as a custom project.
EXTINCTION
Is my target gene silenced at 100%?
The extinction level observed in all established SilenciX cell line is
around 70%, leading to a very low or undetectable expression of
silenced genes. Nevertheless, silencing levels might vary between cell lines. As SilenciX technology knocks-down gene expression
(and not knock-out), a residual expression still remains. It has been
shown that even if the silencing is slight, the loss of protein
function can affect the normal cellular mechanism and the cell acts as
silenced one
What's happened in case of redundant genes?
A loss of function should be observed unless redundant genes take over
functional recovery. Such take over may also be observed in transient
silencing and is not specific to the SilenciX technology. If the
redundancy is known, it might be worth making a single knock-down for
each redundant gene and a double knock-down for the sake of
comparison.
Is there a possibility to switch off the silencing?
Yes, it is feasible 2 months after the transfection.
Nevertheless, we would advise to compare SilenciX cell lines to
control cells.
CONTROL
What is the control for specific silencing?
We supply a standard control cell line with each SilenciX cell line.
The control cell line is established by using a designed standard
control shRNA that has been proven to be inactive. For each catalog
SilenciX cell line puchase, control cell line is included in the parcel.
Can my own validated control siRNA sequence be used for making a
control cell line?
Yes, we will produce it as a custom project.
How do you check the silencing of my gene?
The silencing of target genes is validated by qPCR and we guarantee
>70% extinction at the mRNA level. Data are shown in product
specification sheets. For certain targets, western-blot or immuno-fluorescence
have been performed and showed.
TECHNICAL QUESTIONS
Which protocol for cell culture would you recommend?
Thawing procedure as well as cell culture protocols are described in
the User Manual supplied in your parcel or can be downloaded from our
web site.
Which contaminants do you test in SilenciX cells?
We check the absence of Mycoplasma in each cell line.
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