Generation of 17C-2 iCas9 hiPSCs containing a custom CRISPR library (17C-2-CC) by lentiviral transduction, associated with Fig 2.

(A) (left) Phase-contrast images of lenti-viral transduced 17C-2 hiPSCs before (day 1) and after (day 3) puromycin selection. Volumes of virus-containing supernatant added per well (0 μl, 8 μl and 16 μl; virus titer) are indicated. (+), with puromycin; (-), without puromycin. (right) The number of cells counted at day 3 in each titer. 0 (-) is set as 100%. Titer 8 was selected as the optimized titer for a multiplicity of infection (MOI) of 0.3. (B) Phase-contrast images of 17C-2-CC hiPSCs (left), iMeLCs (middle) or day 5 floating aggregates containing hPGCLCs derived from 17C-2-CC hiPSCs (right). Bars, 200 μm. (C) Distribution of abundances of normalized sgRNA read counts in all screen samples. (D) Comparison of read counts of all sgRNAs in biological replicates of PGCLC(+) screen samples. (E) Normalized read counts of sgRNAs targeting TFAP2C (left) and SOX17 (right) in PGCLC(+) vs. PGCLC(-) populations in screen replicates. Read counts in in PGCLC(-) cells set to 1.0.

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