en Medical Biology Medical Biology مقاله پژوهشی Original Research Article <p style="text-align: justify;"><span style="font-size:16px;"><span style="font-family:Times New Roman;"><strong new="" roman="" style="font-family: " times=""><span style="color:#ffffff;"><span style="background-color:#16a085;">Background and Objective:</span></span>&nbsp;</strong><span style="line-height:200%"><span lang="EN-CA">Medulloblastoma formation is importantly related to granular cell proliferation and differentiation, processes which are under the influence of sonic hedgehog (SHH) signaling. Exons of genes encoding different components of this signaling pathway (e.g., ligands, co-receptor, transcription factors, and target genes) were investigated to identify the proper single guide RNAs for selective targeting of positive regulators of the SHH pathway.</span></span></span></span><br> <span style="font-size:16px;"><span style="font-family:Times New Roman;"><strong new="" roman="" style="font-family: " times=""><span style="color:#ffffff;"><span style="background-color:#16a085;">Materials and Methods:</span></span>&nbsp;</strong><span style="line-height:200%"><span lang="EN-CA">The genomic DNA sequences of corresponding genes of several positive regulators of the SHH pathway (in Homo Sapiens), including <i>SHH</i>, <i>SMO</i>, <i>GLI1, GLI2, MYCN</i>, and <i>MYC</i>,<i> </i>were retrieved from the National Center for Biotechnology Information (NCBI) gene database. Next, the exon sequences of these genes were identified using protospacer adjacent motif (PAM) of NGG and Streptococcus pyogenes Cas9 nuclease and evaluated by CRISPOR software to select the best sgRNA for each target gene.</span></span></span></span><br> <span style="font-size:16px;"><span style="font-family:Times New Roman;"><strong new="" roman="" style="font-family: " times=""><span style="color:#ffffff;"><span style="background-color:#16a085;">Results:</span></span></strong>&nbsp;<span style="line-height:200%"><span lang="EN-CA">The analyses revealed the best sgRNAs for the <i>SHH</i>, <i>SMO</i>, <i>GLI1</i>, <i>GLI2</i>, <i>MYCN</i>, and <i>MYC</i> genes targeted exon 1, -4, -4, -5, -2 and -2, respectively. Proper sgRNAs for targeting each exon in each gene were also identified.</span></span></span></span><br> <span style="font-size:16px;"><span style="font-family:Times New Roman;"><strong new="" roman="" style="font-family: " times=""><span style="color:#ffffff;"><span style="background-color:#16a085;">Conclusion:</span></span>&nbsp;</strong><span style="line-height:200%"><span lang="EN-CA">This study revealed possible specific exonic targets of components of the SHH signaling pathway through designing proper sgRNAs using the CRISPR/Cas9 genome editing approach.</span></span></span></span></p> <p><p style="text-align: justify;"></p></p> CRISPR/Cas9, Sonic Hedgehog, Medulloblastoma, Guide RNA, Genome 594 601 http://journal.zums.ac.ir/browse.php?a_code=A-10-2691-5&slc_lang=en&sid=1 Mehrdad Ghorbanlou mehrdad.ghorbanlou@gmail.com 5200319475328460079158 5200319475328460079158 No Dept. of Anatomy, School of medicine, Iran University of Medical Sciences, Tehran, Iran Hasan Marzban hassan.marzban@umanitoba.ca 5200319475328460079159 5200319475328460079159 No Children’s Hospital Research Institute of Manitoba (CHRIM), College of Medicine, Faculty of Health Sciences, University of Manitoba, Winnipeg, MB, Canada Mehdi Mehdizadeh mehdizadeh.m@iums.ac.ir 5200319475328460079160 5200319475328460079160 Yes Reproductive Sciences and Technology Research Center, School of Medicine, Iran University of Medical Sciences, Tehran, Iran