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. 2022 Nov;20(11):2048-2050.
doi: 10.1111/pbi.13915. Epub 2022 Sep 15.

High-throughput sequencing revealed low-efficacy genome editing using Cas9 RNPs electroporation and single-celled microinjection provided an alternative to deliver CRISPR reagents into Euglena gracilis

Zhenfan Chen  1 , Jiayi Zhu  1 , Zixi Chen  1 , Ming Du  1 , Rao Yao  1 , Wen Fu  1 , Anping Lei  1 , Jiangxin Wang  1
Affiliations

High-throughput sequencing revealed low-efficacy genome editing using Cas9 RNPs electroporation and single-celled microinjection provided an alternative to deliver CRISPR reagents into Euglena gracilis

Zhenfan Chen et al. Plant Biotechnol J. 2022 Nov.
No abstract available

Keywords: Euglena gracilis; CRISPR; Electroporation; Gene editing; Microinjection.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
CRISPR genome editing to target the EgGSL2 gene by electroporation and the crtP1 gene using microinjection to deliver Cas9‐RNPs into E. gracilis. (a) Overview of research status on E. gracilis. The data on the chart are derived from Web of Science and are not strictly proportional to the values of each entry. (b) The cleavage experiment on the partial EgGSL2 gene using RNPs with the sgRNA‐target1 designed by Nomura et al. (c) Overview of electroporation parameters to deliver RNPs into E. gracilis. (d) Detection of mutations in three EG300 replicates at EgGSL2 target sites after electroporation using the T7E I assay at 72 h. (e) Detection of mutant sequences in the EgGSL2 gene after electroporation by high‐throughput sequencing. (f) Detection of the CRISPR genome edited mutant sequences and mutant rate of the EgGSL2 gene after electroporation, by high‐throughput sequencing. (g) Representative images of delivering DAPI stain into an E. gracilis cell by microinjection. (h) Schematic of CRISPR genome editing targeting the crtP1 gene.
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References

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