Extended Data Fig. 1: Generation of CERST mouse lines.
a, Gene targeting of the TIGRE locus. The gRNA cassette and the V1 recorder cassette were targeted into the TIGRE locus of mouse embryonic stem cells (mESCs). The upper diagram shows the wild-type TIGRE locus, the lower shows the donor plasmid used for gene targeting. The probes used for southern blot are shown. Neo, neomycin resistance gene driven by PGK promoter; Frt, recognition site of Flp recombinase; DTA, diphtheria toxin A. b, Southern blot of selected mESC clones. The probes used are indicated in Extended Data Fig. 1a. Successful targeting of clone 3-H03 was confirmed using restriction fragments of different sizes: 19.7 kb for the 5′ probe, 6.3 kb for the 3′ probe, and 11.7 kb for the Neo probe. c, BFP expression was observed in the 3-H03 mESC clone, which was used to generate the TIGRECREST-gRNAs mouse line. d, Gene targeting of Rosa26 locus. The Cas9 cassette and the V2 recorder cassette were targeted into the Rosa26 locus of mouse zygotes. The upper diagram shows the wild-type Rosa26 locus, and the lower shows the donor plasmid used for gene targeting. e, Successful targeting of Rosa26CREST-Cas9 mouse was confirmed through genotyping using the primers indicated in Extended Data Fig. 1d. f, Retrieval of CREST barcodes for individual cells. CREST barcodes are enriched from cDNA libraries of single cells, classified as V1 or V2, and assigned to individual cells by 10X cell barcodes for clonal analysis (see Methods).
