English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Background  β-Thalassemia is an inherited hematological disorder caused by mutations in the human hemoglobin beta ( HBB ) gene that reduce or abrogate β-globin expression. Although lentiviral-mediated expression of β-globin and autologous transplantation is a promising therapeutic approach, the risk of insertional mutagenesis or low transgene expression is apparent. However, targeted gene correction of  HBB  mutations with programmable nucleases such as CRISPR/Cas9, TALENs, and ZFNs with non-viral repair templates ensures a higher safety profile and endogenous expression control.    Methods  We have compared three different gene-editing tools (CRISPR/Cas9, TALENs, and ZFNs) for their targeting efficiency of the  HBB  gene locus. As a proof of concept, we studied the personalized gene-correction therapy for a common β-thalassemia splicing variant  HBB  IVS1–110  using  Cas 9 mRNA and several optimally designed single-stranded oligonucleotide (ssODN) donors in K562 and CD34 +  hematopoietic stem cells (HSCs).    Results  Our results exhibited that indel frequency of CRISPR/Cas9 was superior to TALENs and ZFNs ( P  < 0.0001). Our designed sgRNA targeting the site of  HBB  IVS1–110  mutation showed indels in both K562 cells (up to 77%) and CD34 +  hematopoietic stem cells—HSCs (up to 87%). The absolute quantification by next-generation sequencing showed that up to 8% site-specific insertion of the  Nhe I tag was achieved using  Cas9  mRNA and a chemically modified ssODN in CD34 +  HSCs.    Conclusion  Our approach provides guidance on non-viral gene correction in CD34 +  HSCs using  Cas 9 mRNA and chemically modified ssODN. However, further optimization is needed to increase the homology directed repair (HDR) to attain a real clinical benefit for β-thalassemia.    Electronic supplementary material  The online version of this article (10.1186/s40348-018-0086-1) contains supplementary material, which is available to authorized users.

Gene correction of HBB mutations in CD34+ hematopoietic stem cells using Cas9 mRNA and ssODN donors