Generation of K562 cell line expressing Cas9 endonuclease (CRISPR-associated9)

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

ANSARI F.; NIKOUGOFTAR ZARIF M.; HAMIDIEH A.A.; SHAMSARA M.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: The Scientific Journal of Iranian Blood Transfusion Year:2019 | Volume:16 | Issue:1 Page(s): 32-43

Download Full-Text

Persian Verion

View:

783

Download:

Cites:

Information Journal Paper

Title

Generation of K562 cell line expressing Cas9 endonuclease (CRISPR-associated9)

Author(s)

ANSARI F. | NIKOUGOFTAR ZARIF M. | HAMIDIEH A.A. | SHAMSARA M. | Issue Writer Certificate

Keywords

Cell LineQ2

ElectroporationQ3

Gene EditingQ1

Abstract

Background and Objectives: The genome of Cell Lines is nowadays edited to create disease models and treat them. Of course, the size of the cas9 gene has caused problems like the low efficiency of the CRISPR system. To solve this problem, Cas9 expressing Cell Lines have been generated in which CRISPR RNA should only be transfected to the cell. Materials and Methods: This article is experimental. PGK-PURO / CMV (PPC) fragment was amplified with PCR from pAAVS1-puro-DNR vector and cloned in pTG19-T vector. The PPC fragment from this vector was removed by KpnI and EcoRI enzymes. Also the pCas-Guide-AAVS1 vector was subjected to the same enzymatic cutting and its attachment to the PPC fragment resulted in the production of the pPPC-Cas vector. After optimizing the Electroporation conditions, the pPPC-Cas vector was electroporated into K562 cells and puromycin-resistant cells were selected and Cas9 expression level was evaluated by Real-time PCR. Results: A PCR fragment of 2514 bp was amplified. The vector pPPC-Cas was cloned in two steps. puromycin-resistant transfected cells were selected. Clonal selection was carried out and three colonies with high, medium and low expression level of Cas9 were isolated. Conclusions: The Cas9-expressing K562 cells derived in this study can be applied both for functional genomic researches and design cellular models of human diseases in future.

Cites

No record.

References

No record.

Cite

APA: Copy

ANSARI, F., NIKOUGOFTAR ZARIF, M., HAMIDIEH, A.A., & SHAMSARA, M.. (2019). Generation of K562 cell line expressing Cas9 endonuclease (CRISPR-associated9). THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), 16(1 ), 32-43. SID. https://sid.ir/paper/394002/en

Vancouver: Copy

ANSARI F., NIKOUGOFTAR ZARIF M., HAMIDIEH A.A., SHAMSARA M.. Generation of K562 cell line expressing Cas9 endonuclease (CRISPR-associated9). THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)[Internet]. 2019;16(1 ):32-43. Available from: https://sid.ir/paper/394002/en

IEEE: Copy

F. ANSARI, M. NIKOUGOFTAR ZARIF, A.A. HAMIDIEH, and M. SHAMSARA, “Generation of K562 cell line expressing Cas9 endonuclease (CRISPR-associated9),” THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), vol. 16, no. 1 , pp. 32–43, 2019, [Online]. Available: https://sid.ir/paper/394002/en

Related Journal Papers