GEO Logo
   NCBI > GEO > Accession DisplayHelp Not logged in | LoginHelp
GEO help: Mouse over screen elements for information.
Series GSE236755 Query DataSets for GSE236755
Status Public on Nov 05, 2023
Title Cas-SF01, an engineered Cas12i nuclease is an efficient genome editing tool in mammalian cells and plants
Organism Homo sapiens
Experiment type Other
Summary The type V-I CRISPR-Cas system is becoming increasingly attractive for its potential utility in gene editing. However, natural nucleases often exhibit low efficiency, limiting their application. Here, we utilized structure-guided rational design and combinatorial protein engineering to optimize an uncharacterized Cas12i nuclease, Cas12i3. Accordingly, we developed Cas-SF01, a Cas12i3 variant that exhibits significantly improved gene-editing activity in mammalian cells and plants. Cas-SF01 displays comparable or superior editing performance compared to SpCas9 or recently engineered Cas12 nucleases. Further analysis of PAM recognition showed that Cas-SF01 has an expanded PAM range and effectively recognizes NTTN and noncanonical NATN and TTVN PAMs. Additionally, we identified an amino acid substitution, D876R, that markedly reduced the off-target effect while maintaining high on-target activity, leading to the development of Cas-SF01HiFi (high-fidelity Cas-SF01). Finally, we demonstrated that Cas-SF01 has robust gene-editing activity in both the monocot plant rice and dicot plant pepper. Our results suggest that Cas-SF01 can serve as a robust gene-editing platform with high efficiency and specificity for future genome editing applications across different organisms.
Overall design to investigate the off-target effect of the Cas12i3 nuclease, we established the GUIDE-Seq experiment of two gene targetes in 293T cell.
Contributor(s) Duan Z, Liang Y
Citation(s) 38379787
Submission date Jul 07, 2023
Last update date Mar 07, 2024
Contact name Weilong Yang
Organization name Southern University of Science and Technology
Street address No. 1088 Xueyuan Avenue, Nanshan District
City Shenzhen
State/province Guangdong
ZIP/Postal code 518055
Country China
Platforms (1)
GPL24676 Illumina NovaSeq 6000 (Homo sapiens)
Samples (8)
GSM7575044 293T,3days, rep1_1
GSM7575045 293T,3days, rep1_2
GSM7575046 293T,3days, rep1_3
BioProject PRJNA992370

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
GSE236755_RAW.tar 450.0 Kb (http)(custom) TAR (of TXT)
SRA Run SelectorHelp
Raw data are available in SRA
Processed data provided as supplementary file

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