Design and Construction of Recombinant CRISPR Vector Harboring LRRK2 Gene for Parkinson's Disease

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Samare Gholami A.; SASAN H.A.; HASHEMABADI M.; Ravan H.

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Journal Paper

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Journal: JOURNAL OF CELL & TISSUE Year:2020 | Volume:10 | Issue:4 Page(s): 214-225

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Title

Design and Construction of Recombinant CRISPR Vector Harboring LRRK2 Gene for Parkinson's Disease

Author(s)

Samare Gholami A. | SASAN H.A. | HASHEMABADI M. | Ravan H. | Issue Writer Certificate

Keywords

CRISPRQ1

Parkinson's diseaseQ1

LRRK2 geneQ1

Abstract

Aim: In this study, the alteration of the Leucine-rich repeat kinase 2 (LRRK2) gene, as the most important gene involved in Parkinson's disease, was investigated using CRISPR-Cas editing technology. Material and methods: Cloning the guide molecules of interest was performed using of gRNA CRISPR cloning kit (Catalog No. C8324K, Zaver Zist Azema). Two sets of forward and reversed gRNA oligonucleotides for exon 41 of the LRRK2 gene were designed using the CHOPCHOP CRISPR guide gRNA designing web software. Double-stranded DNA molecules were made according to standard instructions in the laboratory. 20 base-pair DNA segments were used to generate RNACas-9 enzyme complex and then ligated into the Px459 CRISPR eukaryotic expression vector using Fermentas DNA ligase enzyme. Recombinant plasmids were transformed into E. coli DH5a host competent cells using of heat shock method. Results: findings by multiple PCR experiments and also DNA sequencing blast analysis showed successful cloning the segments of interest into CRISPR plasmids. Clear PCR bands were in agreement with expected values and DNA sequencing results showed 100% similarity. Conclusion: According to the results, development of CRISPR vector system may be used as a wide and powerful tool for editing and alteration of many encountered genes in different diseases such as Parkinson's disease.

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APA: Copy

Samare Gholami, A., SASAN, H.A., HASHEMABADI, M., & Ravan, H.. (2020). Design and Construction of Recombinant CRISPR Vector Harboring LRRK2 Gene for Parkinson's Disease. JOURNAL OF CELL & TISSUE, 10(4 ), 214-225. SID. https://sid.ir/paper/364460/en

Vancouver: Copy

Samare Gholami A., SASAN H.A., HASHEMABADI M., Ravan H.. Design and Construction of Recombinant CRISPR Vector Harboring LRRK2 Gene for Parkinson's Disease. JOURNAL OF CELL & TISSUE[Internet]. 2020;10(4 ):214-225. Available from: https://sid.ir/paper/364460/en

IEEE: Copy

A. Samare Gholami, H.A. SASAN, M. HASHEMABADI, and H. Ravan, “Design and Construction of Recombinant CRISPR Vector Harboring LRRK2 Gene for Parkinson's Disease,” JOURNAL OF CELL & TISSUE, vol. 10, no. 4 , pp. 214–225, 2020, [Online]. Available: https://sid.ir/paper/364460/en

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