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Deleting 7Mbp chromosomal region


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#1 Cynthia

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Posted 29 March 2014 - 08:58 AM

Hi,

I would like to delete a 7Mbp region on one chromosome in a myeloma cell line and study the effect on immunoglobulin production. Is there a way to delete such a large region?

Thank,

Cynthia



#2 jerryshelly1

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Posted 29 March 2014 - 11:42 AM

Do you need to delete it completely or can you just disrupt the sequence to hinder production?



#3 Cynthia

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Posted 29 March 2014 - 12:26 PM

Either would be fine. I don't want the genes in the region to be expressed so that I could check if the region is important for a phenotype.



#4 perneseblue

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Posted 30 March 2014 - 06:22 AM

Literature search indicate that people have succeeded in deleting upto 10Mb using Cre-loxP system. I have succeeded in deleting 2Mb using Cre-loxP but failed at 4Mb.

So in my opinion it is possible to delete 7Mb but it will be rather difficult and definitely time consuming. 

 

It would be better to use CRISPR system to knock out the gene of interest. It would be faster.


May your PCR products be long, your protocols short and your boss on holiday




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