Generation of global Spata19 knockout mouse using CRISPR/Cas9 nickase technology

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Zargar Mahsa; Jamshidizad Abbas; Rahim Tayefeh Aidin; Hashemi Ehsan; NAJAFI ALI; SHAMSARA MEHDI; MODARRESSI MOHAMMAD HOSSEIN

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Journal: Koomesh Year:2020 | Volume:22 | Issue:3 (79) Page(s): 380-388

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Title

Generation of global Spata19 knockout mouse using CRISPR/Cas9 nickase technology

Author(s)

Keywords

Knock OutQ1

Spata19Q1

CRISPR/Cas9 NickaseQ1

MouseQ1

SpermQ2

pX335Q2

Abstract

Introduction: Spata19 gene is expressed in developmental stages of testis and some organs, but so far its function has only been examined in the testis. In this study, we provided an effective pathway for the generation of these mice using new CRISPR / Cas9 nickase method while generating Spata19 knockout mice for future studies in other organs. Materials and Methods: CRISPR / Cas9 nickase plasmids were synthesized using pX335 vector and designed gRNAs. So, each plasmid was transformed into E. coli and clonal selection was performed. The plasmids extracted from the bacteria were mixed in equal proportions and injected into the male pronucleus. Correspondingly, the zygotes carrying the plasmid were transferred to Oviduct of Foster mice. After pregnancy extent, born mice were analyzed. Results: We generated global knockout mice using CRISPR/Cas9 Nickase technology. This deletion was examined at three levels of DNA, RNA and protein. Generated 2 nucleotides deletion was confirmed by sequencing at three levels of DNA, RNA and protein. By translating the mutated RNA sequence, it was determined that this deletion could cause premature stop codon. Phenotypic analysis confirmed infertility of male homozygous mice (Spata19-/-). Conclusion: By generating Spata19 knockout mice, we present a protocol for generation of knockout mice. These mice could be applied as the infertile male mice and also for further characterization of Spata19 gene.

Cites

Spata19 Inactivation as a Cause of Oligospermia

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APA: Copy

Zargar, Mahsa, Jamshidizad, Abbas, Rahim Tayefeh, Aidin, Hashemi, Ehsan, NAJAFI, ALI, SHAMSARA, MEHDI, & MODARRESSI, MOHAMMAD HOSSEIN. (2020). Generation of global Spata19 knockout mouse using CRISPR/Cas9 nickase technology. KOOMESH, 22(3 (79) ), 380-388. SID. https://sid.ir/paper/397441/en

Vancouver: Copy

Zargar Mahsa, Jamshidizad Abbas, Rahim Tayefeh Aidin, Hashemi Ehsan, NAJAFI ALI, SHAMSARA MEHDI, MODARRESSI MOHAMMAD HOSSEIN. Generation of global Spata19 knockout mouse using CRISPR/Cas9 nickase technology. KOOMESH[Internet]. 2020;22(3 (79) ):380-388. Available from: https://sid.ir/paper/397441/en

IEEE: Copy

Mahsa Zargar, Abbas Jamshidizad, Aidin Rahim Tayefeh, Ehsan Hashemi, ALI NAJAFI, MEHDI SHAMSARA, and MOHAMMAD HOSSEIN MODARRESSI, “Generation of global Spata19 knockout mouse using CRISPR/Cas9 nickase technology,” KOOMESH, vol. 22, no. 3 (79) , pp. 380–388, 2020, [Online]. Available: https://sid.ir/paper/397441/en

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