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10.1038/srep16031 http://scihub22266oqcxt.onion/10.1038/srep16031 C4642230!4642230 !26558999     free     free     free Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 249.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 249.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 249.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 249.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\26558999 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Sci+Rep 2015 ; 5 (ä): 16031 Nephropedia Template TP {{tp|p=26558999 |t=2015. Controlled delivery of ?-globin-targeting TALENs and CRISPR/Cas9 into mammalian cells for genome editing using microinjection |pdf=|usr=}}{{26558999 }} gab.com Text Controlled delivery of -globin-targeting TALENs and CRISPR/Cas9 into mammalian cells for genome editing using microinjection JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=26558999 #FOAvip Tal-effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR) with CRISPR-associated (Cas) proteins are genome editing tools with unprecedented potential. However, the ability to deliver optimal amounts of these nucleases into mammalian cells with minimal toxicity poses a major challenge. Common delivery approaches are transfection- and viral-based methods; each associated with significant drawbacks. An alternative method for directly delivering genome-editing reagents into single living cells with high efficiency and controlled volume is microinjection. Here, we characterize a glass microcapillary-based injection system and demonstrate controlled co-injection of TALENs or CRISPR/Cas9 together with donor template into single K562 cells for targeting the human ?-globin gene. We quantified nuclease induced insertions and deletions (indels) and found that, with ?-globin-targeting TALENs, similar levels of on- and off-target activity in cells could be achieved by microinjection compared with nucleofection. Furthermore, we observed 11% and 2% homology directed repair in single K562 cells co-injected with a donor template along with CRISPR/Cas9 and TALENs respectively. These results demonstrate that a high level of targeted gene modification can be achieved in human cells using glass-needle microinjection of genome editing reagents. Twit Text FOAVip Controlled delivery of -globin-targeting TALENs and CRISPR/Cas9 into mammalian cells for genome editing using microinjection ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=26558999 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26558999 #FOAvip English Wikipedia <ref>{{Cite pmid|26558999 }}</ref> Controlled delivery of ?-globin-targeting TALENs and CRISPR/Cas9 into mammalian cells for genome editing using microinjection #MMPMID26558999 Cottle RN ; Lee CM ; Archer D ; Bao G Sci Rep 2015[Nov]; 5 (ä): 16031 PMID26558999 show ga Tal-effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR) with CRISPR-associated (Cas) proteins are genome editing tools with unprecedented potential. However, the ability to deliver optimal amounts of these nucleases into mammalian cells with minimal toxicity poses a major challenge. Common delivery approaches are transfection- and viral-based methods; each associated with significant drawbacks. An alternative method for directly delivering genome-editing reagents into single living cells with high efficiency and controlled volume is microinjection. Here, we characterize a glass microcapillary-based injection system and demonstrate controlled co-injection of TALENs or CRISPR/Cas9 together with donor template into single K562 cells for targeting the human ?-globin gene. We quantified nuclease induced insertions and deletions (indels) and found that, with ?-globin-targeting TALENs, similar levels of on- and off-target activity in cells could be achieved by microinjection compared with nucleofection. Furthermore, we observed 11% and 2% homology directed repair in single K562 cells co-injected with a donor template along with CRISPR/Cas9 and TALENs respectively. These results demonstrate that a high level of targeted gene modification can be achieved in human cells using glass-needle microinjection of genome editing reagents. |*CRISPR-Cas Systems [MESH] |*Gene Targeting [MESH] |*Genetic Engineering [MESH] |*Genome [MESH] |*Microinjections/methods [MESH] |Animals [MESH] |Cell Line [MESH] |Endonucleases/*genetics [MESH] |Gene Expression [MESH] |Gene Transfer Techniques [MESH] |Humans [MESH] |INDEL Mutation [MESH] |K562 Cells [MESH] |Mutagenesis, Insertional [MESH] |Single-Cell Analysis [MESH]Controlled delivery of ?-globin-targeting TALENs and CRISPR/Cas9 into (epmc).pdf DeepDyve Pubget Overpricing