Gene Editing via Integrase Enzyme
Department of Biology, Comsats University, Islamabad, Pakistan
Received Date: 03/12/2021; Published Date: 10/01/2022
*Corresponding author: Umair Masood, Department of Biology, Comsats University, Islamabad, Pakistan
Cite this article: Umair Masood*. Gene Editing via Integrase Enzyme.
Targeted integrase enzyme is a most powerful tool for mediating genome alteration with high precision. The gene of the interest is directly catalyze nucleophilic attack of the 3 prime hydroxyl group at the end of processed DNA on a pair of phosphodiaster bond in the targeted DNA or genome of the interest. Integrase gene editing method contain two parts gene of the interest with integrase enzyme and desire genome. The integrase enzyme is start integration the gene of the interest into the desire genome or DNA.
Results via Gel electrophoresis
In order to check that whether gene of interest is integrated or not we can perform a gel electrophoresis. The gel contain a two band A and B.A band is a vector of human insulin vector which can show the negative control and B band is a vector of human insulin but the gene have some sequence mutation [1-5]. We can add a gene of the stranded human insulin by using integrase enzyme and gene can be precisely add to the B band that why B band is in 3.0 and A band is 6.0 which mean that A band is high molecular weight than the B band and a Marker must be 1kb and agarose should be 1% [6-9].
- Varmus H, Brown PO (1989) Retroviruses. In: Berg DE, Howe MM, editors. Mobile DNA. Washington, DC, USA: American Society of Microbiology pp. 53–108.
- Coffin JM, Hughes SH, Varmus HE (1977) Retroviruses.Cold Spring Harbor Laboratory Press, USA.
- Richman DD, Margolis DM, Delaney M, Greene WC, Hazuda D, et al. (2009) The challenge of finding a cure for HIV infection. Science 323(5919):1304–1307.
- Bowerman B, Brown PO, Bishop JM, Varmus HE (1989) A nucleoprotein complex mediates the integration of retroviral DNA. Genes Dev3: 469–478.
- Arnet CM, Haseltine WA (1990) Integration of human immunodeficiency virus type 1 DNA in vitro. Proc. Natl. Acad. Sci. USA, 87: 4164–4168.
- Brown PO, Bowerman B, Varmus HE, Bishop JM (1987) Correct integration of retroviral DNA in vitro. Cell 49: 347–356.
- Yin Z, Craigie R (2010) Modeling the HIV-1 intasome: a prototype view of the target of integrase inhibitors. Viruses 2(12): 2777–2781.
- Lee K, Ambrose Z, Martin TD (2010) Flexible use of nuclear import pathways by HIV-1. Cell Host Microbe 7(3): 221–233.
- Zheng RL, Ghirlando R, Lee MS, Mizuuchi K, Krause M, et al. (2000) Barrier-to-autointegration factor (BAF) bridges DNA in a discrete, higher-order nucleoprotein complex. Proc. Natl Acad. Sci. USA 97(16): 8997–9002.