Restore natural fertility of Kitw/Kitwv mouse with nonobstructive azoospermia through gene editing on SSCs mediated by CRISPR-Cas9

Li, Xiaoyü; Sun, Tie-Cheng; Wang, Xiuxia; Tang, Ji‐Xin; Liu, Yi‐Xun

doi:10.1186/s13287-019-1386-7

Cited by 20 publications

(18 citation statements)

References 24 publications

(30 reference statements)

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“…As discussed later, two groups [9,10] have reported the successful production of GE mice through infection of preimplantation embryos with recombinant adeno-associated viruses (rAAVs) carrying genome-editing components. Recently, spermatogonial stem cells have been found to serve as a promising target for the production of GE mice [11][12][13][14][15]. This technology is called "spermatogonial stem cell-mediated transgenesis", and the schematic representation is shown in Figure 2.…”

Section: Developments In Genome-edited (Ge) Mice Production Technologymentioning

confidence: 99%

“…The sperm cells derived from these transplanted spermatogonial stem cells can fertilize egg cells after intracytoplasmic sperm injection (ICSI). A few research groups have reported success in their attempts to produce GE mice by modifying the genome of spermatogonial stem cells [12][13][14][15]. The embryos resulting from in vitro gene delivery into isolated zygotes/2-cell embryos or ICSI using GE sperm are transferred to the reproductive tracts of pseudo-pregnant female mice to enable their development to full-term.…”

Section: Developments In Genome-edited (Ge) Mice Production Technologymentioning

confidence: 99%

“…In 2015, genome-editing experiments with germline stem cells have been reported to be successful in mice [12][13][14][15]. Wu et al [13] performed in vitro EP of CRISPR/Cas9 and exogenous wild-type 89-bp ssODNs into germline stem cells isolated from mutant mice (Crygc −/− )-carrying a single-nucleotide deletion in the crystalline gamma C (Crygc) gene-suffering from cataract.…”

Section: Gene Delivery To Spermatogonial Stem Cellsmentioning

confidence: 99%

“…Requires ICSI and egg transfer to allow further development of the treated embryos [12][13][14][15][36][37][38][39][40][41][42][43][44][45]…”

Section: Ex Vivomentioning

confidence: 99%

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Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ cells, Embryos, and Fetuses in Mice

Takabayashi

Akasaka

Nakamura

2020

Cells

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The recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) systems that occur in nature as microbial adaptive immune systems are considered an important tool in assessing the function of genes of interest in various biological systems. Thus, development of efficient and simple methods to produce genome-edited (GE) animals would accelerate research in this field. The CRISPR/Cas9 system was initially employed in early embryos, utilizing classical gene delivery methods such as microinjection or electroporation, which required ex vivo handling of zygotes before transfer to recipients. Recently, novel in vivo methods such as genome editing via oviductal nucleic acid delivery (GONAD), improved GONAD (i-GONAD), or transplacental gene delivery for acquiring genome-edited fetuses (TPGD-GEF), which facilitate easy embryo manipulation, have been established. Studies utilizing these techniques employed pregnant female mice for direct introduction of the genome-editing components into the oviduct or were dependent on delivery via tail-vein injection. In mice, embryogenesis occurs within the oviducts and the uterus, which often hampers the genetic manipulation of embryos, especially those at early postimplantation stages (days 6 to 8), owing to a thick surrounding layer of tissue called decidua. In this review, we have surveyed the recent achievements in the production of GE mice and have outlined the advantages and disadvantages of the process. We have also referred to the past achievements in gene delivery to early postimplantation stage embryos and germ cells such as primordial germ cells and spermatogonial stem cells, which will benefit relevant research.

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Section: Developments In Genome-edited (Ge) Mice Production Technologymentioning

confidence: 99%

Section: Developments In Genome-edited (Ge) Mice Production Technologymentioning

confidence: 99%

Section: Gene Delivery To Spermatogonial Stem Cellsmentioning

confidence: 99%

“…Requires ICSI and egg transfer to allow further development of the treated embryos [12][13][14][15][36][37][38][39][40][41][42][43][44][45]…”

Section: Ex Vivomentioning

confidence: 99%

See 2 more Smart Citations

Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ cells, Embryos, and Fetuses in Mice

Takabayashi

Akasaka

Nakamura

2020

Cells

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“…Nonobstructive azoospermia (NOA) is defined as one of the most severe forms of male infertility. Considering that NOA is characterized by the absence of sperm after ejaculation due to failure of spermatogenesis, Li et al [ 58 ] aimed to restore fertility in mouse heterozygous mutant for a c-Kit gene (Kit w /Kit wv ). They first isolated mutant SSCs from one single unilateral testis of a two-week-old specimen propagated in vitro.…”

Section: Alternative Viewmentioning

confidence: 99%

Mini-Review Regarding the Applicability of Genome Editing Techniques Developed for Studying Infertility

et al. 2021

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Infertility is a highly debated topic today. It has been long hypothesized that infertility has an idiopathic cause, but recent studies demonstrated the existence of a genetic substrate. Fortunately, the methods of editing the human genome proven to be revolutionary. Following research conducted, we identified a total of 21 relevant studies; 14 were performed on mice, 5 on zebrafish and 2 on rats. We concluded that over forty-four genes in total are dispensable for fertility in both sexes without affecting host homeostasis. However, there are genes whose loss-of-function induces moderate to severe phenotypic changes in both sexes. There were situations in which the authors reported infertility, exhibited by the experimental model, or other pathologies such as cryptorchidism, cataracts, or reduced motor activity. Overall, zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 are techniques that offer a wide range of possibilities for studying infertility, even to create mutant variants. It can be concluded that ZFNs, TALENs, and CRISPR/Cas9 are crucial tools in biomedical research.

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Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice

Xia,

Wang,

Tan

et al. 2023

Advanced Science

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Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense‐point‐mutation mouse model (LhcgrW495X) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild‐type SLCs are transplanted into LhcgrW495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein‐split PEmax system via lentivirus successfully corrects the mutation in SLCs from LhcgrW495X mice ex vivo. Gene‐corrected SLCs from LhcgrW495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene‐corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in LhcgrW495X mice. Altogether, these results suggest that PE‐based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system.

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Restore natural fertility of Kitw/Kitwv mouse with nonobstructive azoospermia through gene editing on SSCs mediated by CRISPR-Cas9

Cited by 20 publications

References 24 publications

Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ cells, Embryos, and Fetuses in Mice

Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ cells, Embryos, and Fetuses in Mice

Mini-Review Regarding the Applicability of Genome Editing Techniques Developed for Studying Infertility

Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice

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