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SilenciX cell lines: FREQUENTLY ASKED QUESTIONS
The SilenciX cell line is are a ready to use system for studying specific silencing of gene of interest. For each reference a gene has been silenced. For exemple, ERCC1 HeLa SilenciX are cells in which the endonuclease ERCC1 involved in DNA repair has been extinguished.
My target is not listed in your catalogue, how to benefit from SilenciX technology?
If you would like to work in HeLa cells, we will prepare SilenciX as a catalogue product. You would agree that this cell line will be put on the catalogue to benefit from our list price.
We also offer custom preparation in any cell line (including HeLa cells) that will not be made available to anybody else.
What is the delivery time of a SilenciX preparation?
70-90 days maximum to silence a target gene in Human 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?
No, SilenciX is only in Human cells as our vector bringing all benefits contains Human promoters.
Do I need a particular safety handling for the use of SilenciX?
Even if shRNA expressing vectors contain small elements of Epstein-Barr virus, they have not the capability to produce viral particles (unlike adenovirus or lentivirus based plasmids). So you do not need any safety handling.
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 catalogue product or any Human cell line you want as a custom project.
Is my target gene silenced at 100%?
The extinction level observed in all established SilenciX cell line is around 80%, leading to a very low or undetectable expression of silenced genes. Nevertheless, silencing level might vary from a target to another one. As SilenciX technology knock-down gene's 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 till 2 months after the transfection. Nevertheless, we would advise to compare SilenciX cell lines to control cells.
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 catalogue SilenciX reference, 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 gene level. Data are shown in product specification sheets. For certain targets, western-blot or immuno-fluorescence have been performed and showed.
Thawing procedure as well as cell culture protocol 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.
General questions
What is SilenciX cell line?The SilenciX cell line is are a ready to use system for studying specific silencing of gene of interest. For each reference a gene has been silenced. For exemple, ERCC1 HeLa SilenciX are cells in which the endonuclease ERCC1 involved in DNA repair has been extinguished.
My target is not listed in your catalogue, how to benefit from SilenciX technology?
If you would like to work in HeLa cells, we will prepare SilenciX as a catalogue product. You would agree that this cell line will be put on the catalogue to benefit from our list price.
We also offer custom preparation in any cell line (including HeLa cells) that will not be made available to anybody else.
What is the delivery time of a SilenciX preparation?
70-90 days maximum to silence a target gene in Human 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?
No, SilenciX is only in Human cells as our vector bringing all benefits contains Human promoters.
Do I need a particular safety handling for the use of SilenciX?
Even if shRNA expressing vectors contain small elements of Epstein-Barr virus, they have not the capability to produce viral particles (unlike adenovirus or lentivirus based plasmids). So you do not need any safety handling.
Target background
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 catalogue 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 80%, leading to a very low or undetectable expression of silenced genes. Nevertheless, silencing level might vary from a target to another one. As SilenciX technology knock-down gene's 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 till 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 catalogue SilenciX reference, 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 gene 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 protocol 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.
Would you like to have more information?
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