TY - JOUR

T1 - Generation of iTAF1-36-H8.1 and iTAF1-36-H8.2 Human-Induced Pluripotent Stem Cell Lines with HAR Deletion in the CNTN6 Gene by CRISPR/Cas9 Genome-Editing Technology

AU - Chvileva, A. s.

AU - Yunusova, A. m.

AU - Pristyazhnyuk, I. e.

AU - Smirnov, A. v.

AU - Ryzhkova, A. s.

AU - Belokopytova, P. s.

AU - Shnaider, T. a.

N1 - Generation of iTAF1-36-H8.1 and iTAF1-36-H8.2 Human-Induced Pluripotent Stem Cell Lines with HAR Deletion in the CNTN6 Gene by CRISPR/Cas9 Genome-Editing Technology / A. S. Chvileva, A. M. Yunusova, I. E. Pristyazhnyuk [et al.] // Russian Journal of Developmental Biology. – 2024. – Vol. 55, No. 5. – P. 296-306. – DOI 10.1134/S1062360424700267. The study was carried out with financial support from the Russian Science Foundation (RSF), project no. 24-24-00447.

PY - 2025/3/17

Y1 - 2025/3/17

N2 - Many structural features of the human brain emerged during evolution as a result of large-scale changes in the molecular genetic mechanisms that control the development of this organ. One of the key genetic alterations involves mutations in regulatory regions of the human genome, which can change the spatiotemporal expression patterns of early developmental genes. This suggestion is supported by the recent discovery of DNA sequences that evolved rapidly in humans, known as human accelerated regions (HARs). These regions are predominantly noncoding and are located near genes involved in human embryonic development and genes expressed in the central nervous system. Most HARs are involved in the regulation of gene expression associated with human brain development, and mutations in them may be linked to the development of such diseases as schizophrenia, autism spectrum disorder, and Simpson-Golabi-Behmel syndrome. One such gene is CNTN6, in which HAR sequences within introns have been identified and found to be deleted due to structural rearrangements in many patients with neuropsychiatric developmental disorders. To study the role of HARs of the CNTN6 gene in the development of the human nervous system, we generated two human induced pluripotent stem cell (iPSC) lines with a homozygous deletion of one of the HARs in the CNTN6 gene (HARsv2_1748) using CRISPR/Cas9 genome-editing technology. The HARsv2_1748 deletion was confirmed by PCR genotyping and Sanger sequencing. Human iPSC lines have normal karyotype, express markers of pluripotency, and are able to spontaneously differentiate into derivatives of three germ layers.

AB - Many structural features of the human brain emerged during evolution as a result of large-scale changes in the molecular genetic mechanisms that control the development of this organ. One of the key genetic alterations involves mutations in regulatory regions of the human genome, which can change the spatiotemporal expression patterns of early developmental genes. This suggestion is supported by the recent discovery of DNA sequences that evolved rapidly in humans, known as human accelerated regions (HARs). These regions are predominantly noncoding and are located near genes involved in human embryonic development and genes expressed in the central nervous system. Most HARs are involved in the regulation of gene expression associated with human brain development, and mutations in them may be linked to the development of such diseases as schizophrenia, autism spectrum disorder, and Simpson-Golabi-Behmel syndrome. One such gene is CNTN6, in which HAR sequences within introns have been identified and found to be deleted due to structural rearrangements in many patients with neuropsychiatric developmental disorders. To study the role of HARs of the CNTN6 gene in the development of the human nervous system, we generated two human induced pluripotent stem cell (iPSC) lines with a homozygous deletion of one of the HARs in the CNTN6 gene (HARsv2_1748) using CRISPR/Cas9 genome-editing technology. The HARsv2_1748 deletion was confirmed by PCR genotyping and Sanger sequencing. Human iPSC lines have normal karyotype, express markers of pluripotency, and are able to spontaneously differentiate into derivatives of three germ layers.

UR - https://www.elibrary.ru/item.asp?id=80498782

U2 - 10.1134/S1062360424700267

DO - 10.1134/S1062360424700267

M3 - Article

VL - 55

SP - 296

EP - 306

JO - Russian Journal of Developmental Biology

JF - Russian Journal of Developmental Biology

SN - 1062-3604

IS - 5

ER -