Scientific Reports, Volume (12), No (1), Year (2022-3)

Title : ( Transposase-CRISPR mediated targeted integration (TransCRISTI) in the human genome )

Authors: MAHERE REZAZADEBAZAZ , Mohammad Mahdi Ghahramani Seno , Hesam Dehghani ,

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Various methods have been used in targeted gene knock-in applications. CRISPR-based knock-in strategies based on homology-independent repair pathways such as CRISPR HITI have been shown to possess the best efficiency for gene knock-in in mammalian cells. However, these methods suffer from the probability of plasmid backbone insertion at the target site. On the other hand, studies trying to combine the targeting ability of the Cas9 molecule and the excision/integration capacity of the PB transposase have shown random integrations. In this study, we introduce a new homology-independent knock-in strategy, Transposase-CRISPR mediated Targeted Integration (TransCRISTI), that exploits a fusion of Cas9 nuclease and a double mutant piggyBac transposase. In isogenic mammalian cell lines, we show that the TransCRISTI method demonstrates higher efficiency (72%) for site-specific insertions than the CRISPR HITI (44%) strategy. Application of the TransCRISTI method resulted in site-directed integration in 4.13% and 3.69% of the initially transfected population in the human AAVS1and PML loci, respectively, while the CRISPR HITI strategy resulted in site-directed integration in the PML locus in only 0.6% of cells. We also observed lower off-target and random insertions in the TransCRISTI group than the CRISPR HITI group. The TransCRISTI technology represents a great potential for the accurate and high-efficiency knock-in of the desired transposable elements into the predetermined genomic locations.


, CRISPR/Cas system, Cas9, Gene Insertion, piggyBac transposase, TransCRISTI
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author = {REZAZADEBAZAZ, MAHERE and Ghahramani Seno, Mohammad Mahdi and Dehghani, Hesam},
title = {Transposase-CRISPR mediated targeted integration (TransCRISTI) in the human genome},
journal = {Scientific Reports},
year = {2022},
volume = {12},
number = {1},
month = {March},
issn = {2045-2322},
keywords = {CRISPR/Cas system; Cas9; Gene Insertion; piggyBac transposase; TransCRISTI},


%0 Journal Article
%T Transposase-CRISPR mediated targeted integration (TransCRISTI) in the human genome
%A Ghahramani Seno, Mohammad Mahdi
%A Dehghani, Hesam
%J Scientific Reports
%@ 2045-2322
%D 2022