Hossein Jafari1, Sara Hesami1, Mojtaba Safi1, Fatemeh Ghasemi1, Mehdi Banan2. 1. Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Evin, Daneshjoo Blvd., Koodakyar St., Tehran, 1985713834, Iran. 2. Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Evin, Daneshjoo Blvd., Koodakyar St., Tehran, 1985713834, Iran. mbbanan@yahoo.com.
Abstract
OBJECTIVE: To knock-in an EGFP cassette into the γ-globin genes of K562 cells via CRISPR/Cas9, and to assess expression and hydroxyurea (HU)-mediated induction of the targeted EGFP transgene. RESULTS: The EGFP cassettes were specifically knocked into the Gγ gene. EGFP expression was detected in the targeted cell population and isolated clones. Furthermore, EGFP transcript and fluorescence levels were significantly induced following HU-treatment. CONCLUSION: This system is readily utilizable for genome scale studies of cis-acting regulatory elements which are implicated in γ-globin expression or HU-mediated induction.
OBJECTIVE: To knock-in an EGFP cassette into the γ-globin genes of K562 cells via CRISPR/Cas9, and to assess expression and hydroxyurea (HU)-mediated induction of the targeted EGFP transgene. RESULTS: The EGFP cassettes were specifically knocked into the Gγ gene. EGFP expression was detected in the targeted cell population and isolated clones. Furthermore, EGFP transcript and fluorescence levels were significantly induced following HU-treatment. CONCLUSION: This system is readily utilizable for genome scale studies of cis-acting regulatory elements which are implicated in γ-globin expression or HU-mediated induction.
Authors: Svetlana A Smirnikhina; Milyausha I Zaynitdinova; Vasilina A Sergeeva; Alexander V Lavrov Journal: Int J Mol Sci Date: 2022-05-26 Impact factor: 6.208