citations

                    Optimization of a DiCre recombinase system with reduced leakage for conditional genome editing of Cryptosporidium
                

Authors:

Yue Huang, Jinli Li, Shifeng Pei, Heng You, Huimin Liu, Yaqiong Guo, Rui Xu, Na Li, Yaoyu Feng, Lihua Xiao

In:

Source: Parasites & Vectors
Publication Date: (2024)
Issue: :

Research Area:

Parasitology

Cells used in publication:

Cryptosporidium parvum: Species:; Tissue Origin:

Platform:

4D-Nucleofector® X-Unit

Experiment

Sporozoites were harvested from C. parvum oocysts that were excysted using bleach and sodium taurocholate treatment as described previously [22]. A total of 5?×?10⁷ sporozoites were resuspended in SF buffer (Lonza, Basel, Switzerland) containing the donor and CRISPR/Cas9 plasmids (50 µg each in 100 µl). Electroporation was performed using the EH100 program on an AMAXA 4D-Nucleofector System (Lonza, Basel, Switzerland) as described previously

Abstract

The dimerizable Cre recombinase system (DiCre) exhibits increased leaky activity in Cryptosporidium, leading to unintended gene editing in the absence of induction. Therefore, optimization of the current DiCre technique is necessary for functional studies of essential Cryptosporidium genes.

Open in PubMed