Hi Siddalingaprasad,

First, you can confirm that your gene editing was sucessfull by running a mismatch cleavage assay using Surveyor nuclease or else.

Then, you can proceed to clonal expension using a limit dilution assay (dilution to 0,5 cells per well) in a 96 well plate. Then, as the cell grows, you transfert them to 48 wells, etc. 

As soon as you have enough cells for a PCR assay, you can extract DNA and run a PCR overlapping your region of interest. Then proceed to Sanger sequencing or run . This way, you will identify the clones that were edited sucessfully. You can then confirm that they are GS deficient by stopping supplementation or western blot. 

Good luck,

Nicolas

http://blog.addgene.org/crispr-101-validating-your-genome-edit

 Thank you for the reply Nicolas Tremblay

I will work on this one

this is a nice suggestion

thank you

Siddalingaprasad

Hi Prasad

In continuation to Nicolas Tremblay suggestions, I also recommend you sequence the transcript also. That also gives good insights and others orthogonal method too.

Good luck