Hi Arturo,

I've seen many different approaches but the most common is one of the 2 inducible/tamoxifen dependent CX3CR1-CRE lines. I'd try i.p. injections with tamoxifen (9mg/40g) dissolved in peanut oil (20mg/ml) using a 28 gauge

insulin syringe. An animal weighing 25g will receive 281ul of tamoxifen solution per injection. Tamoxifen from sigma catalog number is T5648. This is adapted from a protocol from Maiken Nedergaard and/or Petr Trvdik's labs through Nathan Anthony Smith. They have used the Gan/Littman (NYU) mice, and injected mice daily for 5 days for cKOs I believe. For turning on reporter or calcium imaging constructs, a single injection is usually used..

Also, I recently came across this paper, they do exactly what you're trying to do: delete TSC1 from microglia in adult mice.

https://www.cell.com/cell-reports/abstract/S2211-1247(18)30159-1

They use the GENSAT line (from MMRRC) and inject tamoxifen daily for 5 days:

"Tamoxifen (Sigma‐Aldrich, Cat#T5648) was dissolved in corn oil (Sigma‐Aldrich, Cat#C8267) at a concentration of 20 mg/ml by shaking overnight at 37°C. Dissolved tamoxifen was stored at 4°C for the duration of injections. Cx3cr1-CreERT2+/- and Cx3cr1-CreERT2+/-TSC1f/f mice at 8-10-week-old of age were injected (i.p) with tamoxifen daily at 100 mg/kg body weight for 5 consecutive days."

Thank you so much Alberto!!!

I really appreciate it!!! Extremely helpful and clear!!!

I will take a look at those papers and references.

Best,

-Arturo-

Huyy.. interesante pregunta y muy buen experimento