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Establishing a pathological basis for androgen replacement therapy requires identifying well-defined disorders of the hypothalamus, pituitary or testis which have a known and clearly defined pathological basis

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FACS-based CRISPR–Cas9 SMARCA2 stability reporter screens The SMARCA2 BD stability reporter FACS-based CRISPR/Cas9 screen, library preparation, next-generation sequencing and data analysis was performed as previously described (53) In brief, doxycycline-inducible Cas9 KBM7 cells stably expressing SMARCA2 BD-mTagBFP–P2A–mCherry were transduced in the presence of 8μg ml -1 polybrene with a ubiquitin-proteasome-system-focused (UPS) sgRNA library targeting 1301 ubiquitin-associated human genes with 6 sgRNAs per gene at a multiplicity of infection of 0.15 and over 1000x library representation (26) [Supplementary Table 2]

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