Figure 2.

In vitro production and CRISPR-Cas9 microinjections of MCM embryos. (A) Timeline of oocyte retrieval and early embryo manipulations. Oocytes were retrieved laparoscopically between 38 and 40 h post-treatment of hCG and then cultured for ~ 4–5 h to allow for in vitro maturation (IVM) of oocytes. Following IVM, mature oocytes were fertilized by ICSI and incubated for ~ 5–7 h prior to embryo microinjection with CRISPR-Cas9 constructs (Cas9 alone or CCR5 RNP), or alternatively, embryos were cultured as control. Representative images of (B) MII MCM oocytes and (C) embryonic development of CCR5 RNP, Cas9 alone and control embryos.