GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
Series GSE136393 Query DataSets for GSE136393
Status Public on Oct 22, 2019
Title Retroelement insertion in a CRISPR/Cas9 editing site in the early embryo intensifies genetic mosaicism
Organism Mus musculus
Experiment type Genome variation profiling by high throughput sequencing
Summary Continued CRISPR/Cas9-mediated editing activity that allows differential and asynchronous modification of alleles in successive cell generations expands allelic complexity. To understand the earliest events during CRISPR/Cas9 editing and the allelic selection among the progeny of subsequent cell divisions, we inspected in detail the genotypes of 4- and 8-cell embryos and embryonic stem cells (ESCs) after microinjection of a CRISPR toolkit into the zygotes. We found a higher editing frequency in 8-cell embryos than in 4-cell embryos, indicating that the CRISPR/Cas9 activity persisted through the 8-cell stage. Analysis of a CRISPR/Cas9 transgenic founder mouse revealed that four different alleles were present in its organs in different combinations and that its germline included three different mutant alleles, as shown by the genotypes of the pups. The indel depth, which measured the extent of indels at the sequence level within single embryos, decreased significantly as the embryos advanced to form ESCs, suggesting that exclusion of fatal indels occurred in the subsequent cell generations. Interestingly, we discovered that the CRISPR sites frequently contained introduced retroelement sequences and that this occurred preferentially with certain classes of retroelements. Therefore, in addition to CRISPR/Cas9’s innate mechanism of separate, differential enzymatic modifications of alleles, the frequent retroelement insertions that occur in early mouse embryos during CRISPR/Cas9 editing further expand the allelic diversity and mosaicism in the resulting transgenic founders.
Overall design Genotyping of CRISPR/Cas9 modified genomic regions by the highthrouput sequencing technology
Contributor(s) Min B, Jeon J
Citation(s) 31781562
Submission date Aug 27, 2019
Last update date Dec 05, 2019
Contact name Byungkuk Min
Organization name KRIBB
Department Development and Differentiation Research Center
Street address 125 Gwahak-ro, Yuseong-gu
City Daejeon
ZIP/Postal code 34141
Country South Korea
Platforms (1)
GPL16417 Illumina MiSeq (Mus musculus)
Samples (41)
GSM4047903 Brca2.4cell.13
GSM4047904 Brca2.4cell.14
GSM4047905 Brca2.6cell.12
BioProject PRJNA562464
SRA SRP219386

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE136393_INDEL_frequencies.xlsx 25.1 Kb (ftp)(http) XLSX
SRA Run SelectorHelp
Raw data are available in SRA
Processed data are available on Series record

| NLM | NIH | GEO Help | Disclaimer | Accessibility |
NCBI Home NCBI Search NCBI SiteMap