Figure 2.
FACS Purification Increases the Speed and Yield of Isogenic Derivation
(A) Post-selection sorting of double-positive biallelic edited cells for SNCAe2 and SNCAe3 using independent sgRNAs. FACS plots are represented with 2% contour lines. For SNCAe3 sgRNA-636, a dotplot is included to show the distribution of 1.2%. Diagram of knockin population types is shown (right).
(B) Yield-purity and purity-purity sorting strategies permit the generation of a homogeneous biallelic knockin population. Diagram of purification population types is shown (right).
(C) Representative post-sorting of single cells for SNCA polyclone. Single-cell gating structures yields high-purity biallelic edited cells. Scale bar, 50 μm.
(D) Representative post-sorting culture for biallelic EGFPpos/dTOMATOposSNCA polyclone. Scale bar, 200 μm.
(E) Sanger sequencing chromatogram of SNCAe2 p.A30P polyclone 632 knockin (KI).
(F) Sanger sequencing chromatogram of SNCAe3 p.A53T polyclone 636 knockin.
(G) Analysis of the polyclone composition as in Figure S3.