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Journal: Nature Communications
Article Title: Search-and-remove genome editing allows selection of cells by DNA sequence
doi: 10.1038/s41467-025-66896-1
Figure Lengend Snippet: a Strategy to apply SNIPE to kill cancer cells based on cancer specific mutations. Wild type cells are colored in gray, cells with cancer mutations in purple and dying cells in red. b Cell growth curves of human 409B2 iPSCs measured by resazurin assay (upper panel). The lower panel shows NGS read frequency for cancer mutation (purple) and wild type cells (gray) in a mixed population over time. c Killing of TP53 knock-out 409B2 cells by SNIPE. Different amounts of TP53 knock-out cells (purple) were mixed with wild type cells (gray). Indels are colored in blue and wild type with indels in light purple. d Cell survival after selection by SNIPE related to ( c ). e K562 cells carry a Philadelphia chromosome, characterized by a reciprocal translocation of chromosome 9 and 22 resulting in the fusion oncogene BCR-ABL1 . f The BCR-ABL fusion generates a new gRNA recognition site in K562 cells. g Cell survival of THP-1 and K562 cells after targeting the BCR-ABL junction. The label “Cas9 cut” indicates CRISPR targeting without SNIPE. h Agarose gel of PCR products of healthy chromosome 22 (297 bp, gray) and Philadelphia chromosome (267 bp, purple). i Quantification of band intensity for K562 cells of ( h ). j Cell survival of heterozygous cancer cell lines RD-ES, HT-29 and HuCC-T1 after targeting the disease allele with SNIPE. k NGS read frequency of cancer locus alleles in RD-ES, HT-29 and HuCC-T1 for mock and SNIPE edit. The cancer mutation allele is colored in purple, wild type in grey, indels in blue and wild type with indels in light purple. Independent biological replicates were performed ( n = 3 for b , c , d , g , h , i , j , k , n = 2 for RDES in j and k) and are depicted as dots for the cancer mutation allele sequence reads and cell survival. Error bars show the s.e.m.
Article Snippet: Human intrahepatic cholangiocarcinoma cells HuCC-T1 (Cytion, catalog no. 300469), the
Techniques: Resazurin Assay, Mutagenesis, Knock-Out, Selection, Translocation Assay, CRISPR, Agarose Gel Electrophoresis, Sequencing