I used CRISPR to eliminate special gene in the mice. know I have single and double KO for male and also female.I would like to know in order to prove the effect of lacking this gene on male fertility. how many male should be used and how many female should be mate with the KO male?
Hi Marzieh,
I do not think that there are set rules for testing this because infertility can be due to ability of male to mate (no vaginal plug in fertile female) or ability of sperm to fertilize in vivo (sperm defect) or sperm motility problem and so on. You need to figure out what is the step causing infertility.
If I were you I will start mating 8-10 wks old knock out male with 2 females (6 cages) and check plugs in the morning for 10 days, than remove the males from females, place each positive plugged female in a separate cage. Look if females become pregnant, Sometime you miss plug, just look if any becomes pregnant from all fertile females (do not use het females for breeding).
This is the beginning= from there you may get some clue which way to move forward- You need to read a large number of male infertility papers in knockout and look for all possibilities. Sometime it could be very simple or sometime you need to go very deep in many directions.
You have also some idea about the gene you knocked out its function and its likelihood to interfere with reproductive function,
Best of luck,
Subhash
PS: for example we reported infertility in KOs: Hrb, BubR1, Cx43, HexB genes mentioned in my profile, that is tip of an iceberg