2018 / 02

Next Paper
DOI: 10.24075/brsmu.2018.022

ORIGINAL RESEARCH

Experimental approaches to the target editing of the CFTR gene using CRISPR-Cas9

Smirnikhina SA1, Anuchina AA1, Kochergin-Nikitsky KS1, Adilgereeva EP1, Yakushina VD1, Lavrov AV1,2
About authors

1 Laboratory of Mutagenesis,
Research Centre for Medical Genetics, Moscow

2 Department of Molecular and Cellular Genetics, Biomedical Faculty,
Pirogov Russian National Research Medical University, Moscow

Correspondence should be addressed: Svetlana Smirnikhina
Moskvorechie 1, Moscow, 115522; moc.liamg@sanihkinrims

About paper

Funding: the section Editing of the CFTR locus was supported by the grant of the Russian Science Foundation (Agreement 17-75-20095), the sections Increasing the expression of guide RNAs and Improving the efficacy of CFTR locus editing were supported by the Russian Academy of Sciences and the state assignment of FASO Russia.

Received: 2018-03-15 Accepted: 2018-03-20 Published online: 2018-07-04
|
Fig. 1. Maps of synthetic plasmids used for F508del mutation editing in the CFTR gene
Fig 2. sgRNAs for the CFTR locus used in this study
Fig. 3. Efficacy of CFTR and EGFP editing in HEK293T cells 48–72 hours after transfection. The results are represented as a mean and a standard error of the mean
Fig. 4. Comparison of CFTR editing efficacy in HEK293T cells. The results are represented as a mean
Fig. 5. Efficacy of CFTRCFTR and EGFP editing in HEK293T cells using sgRNA expressed from the standard U6 and the hybrid U6-tRNAgln (plasmid +HP) promoters. The results are represented as a mean
Fig. 6. Comparison of CFTR editing efficacy in HEK293T cells using modified sgRNAs. The results are represented as a mean
Table. Comparison of CFTR editing efficacy using transient hypothermia of HEK293T cells