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Supplemental Material for Sizova et al., 2021

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posted on 06.04.2021, 16:55 by Irina Sizova, Simon Kelterborn, Valeriy Verbenko, Suneel Kateriya, Peter Hegemann

Figure S1 contains growth curves of the parental Chlamydomonas reinhardtii CC-125 strain and mutants in TAP liquid media. Figure S2 contains data on sensitivity of Chlamydomonas reinhardtii mutants to zeocin treatment. Figure S3 contains schematic representation of the mut-aphVIII repair assay in the Chlamydomonas CC3403-D5 strain. Figure S4 contains data on homology directed mut-aphVIII repair frequencies upon CRISPR/Cas9-induced double-strand breaks through oligonucleotides and a plasmid DNA in the parental Chlamydomonas CC3403-D5 strain. Figure S5 contains data on efficiency of the homology directed mut-aphVIII repair upon CRISPR/SpCas9-induced double-strand breaks through oligonucleotides in the parental Chlamydomonas CC3403-D5 strain and its KU80 deficient mutants. Figure S6 contains data on efficiency of the homology directed mut-aphVIII repair upon CRISPR/LbCas12a-induced double-strand breaks through oligonucleotides or a plasmid DNA in the parental Chlamydomonas CC3403-D5 strain and its POLQ deficient mutant. Figure S7 contains data on blue-green screen and analysis of Chlamydomonas colonies containing targeted insertions in the SNRK2.2 gene. Table S1 contains a list of Chlamydomonas strains used in the work including sequences of insertions inactivating the KU80, KU70, POLQ genes. Table S2 contains sequences of mutations generated in the APT gene. Table S3 contains sequences of donor DNA used in the work. Table S4 contains sequences of oligonucleotides used for screening and sequencing. Table S5 contains protospacer sequences.

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Chlamydomonas POLQ is necessary for CRISPR/Cas9-mediated gene targeting

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