Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice.
Human CD34+ hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate "humanized mice". To further extend the range of applications of these humanized mice, we developed a protoco...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2023-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0287052 |
| _version_ | 1797788277061713920 |
|---|---|
| author | Daniel A Kuppers Jonathan Linton Sergio Ortiz Espinosa Kelly M McKenna Anthony Rongvaux Patrick J Paddison |
| author_facet | Daniel A Kuppers Jonathan Linton Sergio Ortiz Espinosa Kelly M McKenna Anthony Rongvaux Patrick J Paddison |
| author_sort | Daniel A Kuppers |
| collection | DOAJ |
| description | Human CD34+ hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate "humanized mice". To further extend the range of applications of these humanized mice, we developed a protocol to efficiently edit the genomes of human CD34+ HSPCs before transplantation. In the past, manipulating HSPCs has been complicated by the fact that they are inherently difficult to transduce with lentivectors, and rapidly lose their stemness and engraftment potential during in vitro culture. However, with optimized nucleofection of sgRNA:Cas9 ribonucleoprotein complexes, we are now able to edit a candidate gene in CD34+ HSPCs with almost 100% efficiency, and transplant these modified cells in immunodeficient mice with high engraftment levels and multilineage hematopoietic differentiation. The result is a humanized mouse from which we knocked out a gene of interest from their human immune system. |
| first_indexed | 2024-03-13T01:34:14Z |
| format | Article |
| id | doaj.art-c40eaaf9412747dea3045397d9b776a9 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-03-13T01:34:14Z |
| publishDate | 2023-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-c40eaaf9412747dea3045397d9b776a92023-07-04T05:32:01ZengPublic Library of Science (PLoS)PLoS ONE1932-62032023-01-01186e028705210.1371/journal.pone.0287052Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice.Daniel A KuppersJonathan LintonSergio Ortiz EspinosaKelly M McKennaAnthony RongvauxPatrick J PaddisonHuman CD34+ hematopoietic stem and progenitor cells (HSPCs) are a standard source of cells for clinical HSC transplantations as well as experimental xenotransplantation to generate "humanized mice". To further extend the range of applications of these humanized mice, we developed a protocol to efficiently edit the genomes of human CD34+ HSPCs before transplantation. In the past, manipulating HSPCs has been complicated by the fact that they are inherently difficult to transduce with lentivectors, and rapidly lose their stemness and engraftment potential during in vitro culture. However, with optimized nucleofection of sgRNA:Cas9 ribonucleoprotein complexes, we are now able to edit a candidate gene in CD34+ HSPCs with almost 100% efficiency, and transplant these modified cells in immunodeficient mice with high engraftment levels and multilineage hematopoietic differentiation. The result is a humanized mouse from which we knocked out a gene of interest from their human immune system.https://doi.org/10.1371/journal.pone.0287052 |
| spellingShingle | Daniel A Kuppers Jonathan Linton Sergio Ortiz Espinosa Kelly M McKenna Anthony Rongvaux Patrick J Paddison Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. PLoS ONE |
| title | Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. |
| title_full | Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. |
| title_fullStr | Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. |
| title_full_unstemmed | Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. |
| title_short | Gene knock-outs in human CD34+ hematopoietic stem and progenitor cells and in the human immune system of mice. |
| title_sort | gene knock outs in human cd34 hematopoietic stem and progenitor cells and in the human immune system of mice |
| url | https://doi.org/10.1371/journal.pone.0287052 |
| work_keys_str_mv | AT danielakuppers geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice AT jonathanlinton geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice AT sergioortizespinosa geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice AT kellymmckenna geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice AT anthonyrongvaux geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice AT patrickjpaddison geneknockoutsinhumancd34hematopoieticstemandprogenitorcellsandinthehumanimmunesystemofmice |