Year: 2019
Type: letter
Abstract: Two papers, by Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus,2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar in the October issue of Molecular Therapy claim that homologous recombination (HR)-based genome editing by Clade F adeno-associated virus (AAV) is inefficient and virtually undetectable in the absence of double-stranded DNA breaks. These claims contradict our findings that Clade F AAVs mediate HR efficiently in primary cells and in vivo in the absence of double-stranded DNA breaks induced by exogenous nucleases.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar We showed that chromosomal insertion of donor sequences by hematopoietic stem cell-derived AAV (AAVHSC) vectors required flanking homology arms and was BRCA2-dependent. Since BRCA2 is an essential mediator of HR,4Prakash R. Zhang Y. Feng W. Jasin M. Homologous recombination and human health: the roles of BRCA1, BRCA2, and associated proteins.Cold Spring Harb. Perspect. Biol. 2015; 7: a016600Crossref PubMed Scopus (479) Google Scholar we concluded that Clade F AAV-mediated editing was accomplished by mediated HR. Importantly, we showed that high MOIs were required to observe augmented HR by Clade F AAV. AAVHSC-mediated HR-based genome editing was found to occur efficiently in vivo and was persistent. Neither Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar nor Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar actually replicated the studies described in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar There were significant discrepancies between our studies and those of Rogers at al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus.2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar These included the MOIs, AAV capsids, transduction, culture conditions, and vector quality. Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar used donor vectors encoding a promoter containing a GFP gene, in contrast to the promoterless editing vectors used in our studies. Both papers only tested in vitro editing with no actual sequence confirmation or in vivo evaluation. Due to these critical discrepancies between their studies and ours, we find their conclusions regarding the ability of Clade F AAV to mediate HR to be flawed. The most significant difference between Rogers et al.,1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar Dudek and Porteus,2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar and Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar lies in the MOIs. In Smith et al.,3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar we clearly showed that efficient in vitro HR-based editing in the absence of nuclease-induced double-stranded DNA breaks was dependent upon the use of high MOIs. 150,000 vector genomes (vg)/cell or higher were required efficient HR. our studies were at 150,000 or L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar In Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar used and Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar used an and that required efficient HR in absence of is that to observe editing in the absence of at these MOIs. In their studies replicated the in Smith et al.,3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar clearly showed that to no HR was at these MOIs with AAV significant of at higher MOIs. Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar showed at with their Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar were to use to regarding the of their vectors and the of high to conclusions regarding the editing of their vectors and regarding the of their The by Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar is in contrast to our in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google we no difference in the of and cells a of MOIs to Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar used a and of their AAV6 vectors with their Clade F AAV6 a in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar or Dudek and Porteus,2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar is editing vectors to to mediate HR. These high of AAV genomes recombination to and H. Ul-Hasan T. the of in the of adeno-associated Ther. PubMed Scopus Google Scholar is the of high editing of AAV L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar chromosomal insertion of AAV donor sequences by of double-stranded DNA breaks by G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar or and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Both used and of we used Clade F AAVs L.J. Ul-Hasan T. M. and of human stem cell-derived Ther. Full Text Full Text PDF PubMed Scopus Google Scholar with and L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar that is to and that an at et L.J. Ul-Hasan T. M. and of human stem cell-derived Ther. Full Text Full Text PDF PubMed Scopus Google Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar by and only a is the of the AAV tested in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar These vectors that were tested in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar and be used to or our Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar and the AAV Clade F with these Since and were used in Smith et al.,3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar to use these vectors to the studies in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Both to the of a at is that to roles in and Full Text Full Text PDF PubMed Scopus Google J. Biol. the of to human PubMed Scopus Google Scholar and be Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar the of the in to in the L.J. Ul-Hasan T. M. and of human stem cell-derived Ther. Full Text Full Text PDF PubMed Scopus Google Scholar that the of and the of of et L.J. Ul-Hasan T. M. and of human stem cell-derived Ther. Full Text Full Text PDF PubMed Scopus Google a these in HR. Since the of in HR is no the AAV vectors used in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar were with virus we of editing studies with AAV vectors and M. M. M. M. T. et of a of Clade F of and the Ther. 2019; 27: Scholar AAV vectors were of and were Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar the of a a virus in HR and of Since AAV vectors tested in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar were and in an is that a in HR only in Clade F AAV. that were the of HR the only Clade F Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar the of to recombination or to AAV in the of the in we clearly showed that BRCA2 was required L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar recombination was in the absence of BRCA2, efficient L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Since no AAVHSC-mediated HR was in by Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar regarding the of the vector of AAV vectors with virus in or be L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google L.J. Ul-Hasan T. M. and of human stem cell-derived Ther. Full Text Full Text PDF PubMed Scopus Google Scholar Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar a of in vitro a of MOIs to the absence of of the In Smith et al.,3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar the editing were with a high of The editing vectors were to promoterless of that be the chromosomal and sequence were included to of the that AAV genomes the was in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus.2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar The absence of in homology arms that the to L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar In Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar only used donor with donor vectors were that occur vectors Both studies GFP in of cells with the that at with no to In Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google we showed that the vector genome in was higher in cells with Clade F by Clade Clade and Clade we of HR by Clade F AAV. In Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar vector genome vector to the and in the in to GFP with vector genome is a between these be GFP and vector genomes were showed with high of GFP and vector genome only in the of and no or vector genome were with and et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Since their AAV6 was and by a their and these the of their vector and or cells at high MOIs. of at MOIs and AAV6 that their AAV6 vector were of higher their and and The culture cells used in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar were those used in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar We that AAV of cells is H. W. of human hematopoietic stem cells adeno-associated virus Ther. PubMed Scopus Google G. of adeno-associated virus vectors in human hematopoietic cells PubMed Google H. J. of human hematopoietic stem cells of in Natl. Acad. Sci. USA. PubMed Scopus Google Scholar and transduction, cells and in culture at high AAV transduction, we that is H. W. of human hematopoietic stem cells adeno-associated virus Ther. PubMed Scopus Google G. of adeno-associated virus vectors in human hematopoietic cells PubMed Google H. J. of human hematopoietic stem cells of in Natl. Acad. Sci. USA. PubMed Scopus Google Scholar to that used in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar their and studies used higher were used in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar Neither of The of sequence confirmation of editing in studies is Dudek and Porteus2Dudek A.M. Porteus M.H. 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PubMed Scopus Google Scholar is associated with of and M. J. is required the Biol. PubMed Scopus Google J. M. is to of cells in to DNA Biol. PubMed Scopus Google Scholar In culture in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus,2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar cells containing AAV genomes that cells in a of the of cells be and is actually L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar AAV at in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus,2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar an of AAV genomes is HR or insertion of or the to of and is that in vitro with AAV vectors these the of AAV vectors is in Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar nor Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar in vivo of In in Smith et al.3Smith L.J. Wright J. Clark G. Ul-Hasan T. Jin X. Fong A. Chandra M. St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google in to in vitro we showed in vivo editing by of the by and and sequence we the ability of AAV6 to cells or In we AAV6 of human hematopoietic stem cells in vitro in Chandra M. 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St Martin T. Rubin H. Knowlton D. et al.Stem cell-derived clade F AAVs mediate high-efficiency homologous recombination-based genome editing.Proc. Natl. Acad. Sci. USA. 2018; 115: E7379-E7388Crossref PubMed Scopus (23) Google Scholar et al.,1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google we that their use of MOIs is the primary their to observe augmented HR by Clade F their use of AAV vectors that were at their ability to observe augmented HR by Clade F AAV. in to observe HR-based editing by Clade F is critical to the of and use of AAV use AAV vectors with and cells that to be efficiently by AAV. HR by Clade F AAVs high MOIs. in the use of high AAV The of Rogers et al.1Rogers G.L. Chen H.-Y. Morales H. Cannon P.M. Homologous recombination-based genome editing by clade F AAVs is inefficient in the absence of a targeted DNA break.Mol. Ther. 2019; 27: 1726-1736Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar and Dudek and Porteus2Dudek A.M. Porteus M.H. AAV6 is superior to Clade F AAVs in stimulating homologous recombination-based genome editing in human HSPCs.Mol. Ther. 2019; 27: 1701-1705Abstract Full Text Full Text PDF PubMed Scopus (7) Google Scholar to cells at 150,000 or higher in the absence of or to our studies clearly their ability to conclusions regarding the HR of Clade F we to to the and use in is of and an in
Source: Molecular Therapy
Author: Saswati Chatterjee
Institution: Beckman Research Institute
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $3,500
OpenAlex ID: W2994233443