Optimising genetic transformation of Trypanosoma cruzi using Hydroxyurea (cas 127-07-1)-induced cell-cycle synchronisation
-
Add time:09/27/2019 Source:sciencedirect.com
The limited flexibility and time-consuming nature of the genetic manipulation procedures applicable to Trypanosoma cruzi continue to restrict the functional dissection of this parasite. We hypothesised that transformation efficiency could be enhanced if electroporation was timed to coincide with DNA replication. To test this, we generated epimastigote cultures enriched at the G1/S boundary using hydroxyurea-induced cell-cycle synchronisation, and then electroporated parasites at various time points after release from the cell-cycle block. We found a significant increase in transformation efficiency, with both episomal and integrative constructs, when cultures were electroporated 1 h after hydroxyurea removal. It was possible to generate genetically modified populations in less than 2 weeks, compared to the normal 4–6 weeks, with a 5 to 8-fold increase in the number of stably transformed clones. This straightforward optimisation step can be widely applied and should help streamline functional studies in T. cruzi.
We also recommend Trading Suppliers and Manufacturers of Hydroxyurea (cas 127-07-1). Pls Click Website Link as below: cas 127-07-1 suppliers
Prev:p53 mediates Hydroxyurea (cas 127-07-1) resistance in aneuploid cells of colon cancer
Next:Acute Hydroxyurea (cas 127-07-1) treatment reduces tubular damage following bilateral ischemia-reperfusion injury in a mouse model of sickle cell disease) - 【Back】【Close 】【Print】【Add to favorite 】
- Related Information
- Clinical studyCutaneous ulcers associated with Hydroxyurea (cas 127-07-1) therapy09/30/2019
- Real-life experience with Hydroxyurea (cas 127-07-1) in sickle cell disease: A multicenter study in a cohort of patients with heterogeneous descent09/29/2019
- Case StudyMalleolar Ulceration Induced by Hydroxyurea (cas 127-07-1) Therapy for Chronic Eosinophila09/28/2019
- Acute Hydroxyurea (cas 127-07-1) treatment reduces tubular damage following bilateral ischemia-reperfusion injury in a mouse model of sickle cell disease10/01/2019
- p53 mediates Hydroxyurea (cas 127-07-1) resistance in aneuploid cells of colon cancer09/26/2019
- Hydroxyurea (cas 127-07-1): Analytical techniques and quantitative analysis09/25/2019
- Hydroxyurea (cas 127-07-1) embryotoxicity is enhanced in P53-deficient mice09/24/2019
- Beliefs about Hydroxyurea (cas 127-07-1) in youth with sickle cell disease09/10/2019
- Original ArticlesTargeted Hydroxyurea (cas 127-07-1) Education after an Emergency Department Visit Increases Hydroxyurea (cas 127-07-1) Use in Children with Sickle Cell Anemia09/09/2019