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10.1186/s40694-015-0015-1 http://scihub22266oqcxt.onion/10.1186/s40694-015-0015-1 C5611662!5611662!28955455     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Fungal+Biol+Biotechnol 2015 ; 2 (ä): ä Nephropedia Template TP {{tp|p=28955455|t=2015. Efficient gene editing in Neurospora crassa with CRISPR technology |pdf=|usr=}}{{28955455}} gab.com Text Efficient gene editing in Neurospora crassa with CRISPR technology JO: Fungal Biol Biotechnol http://www.kidney.de/pget.php?&tw=1&pmid=28955455 #FOAvip Background: Efficient gene editing is a critical tool for investigating molecular mechanisms of cellular processes and engineering organisms for numerous purposes ranging from biotechnology to medicine. Recently developed RNA-guided CRISPR/Cas9 technology has been used for efficient gene editing in various organisms, but has not been tested in a model filamentous fungus, Neurospora crassa. Findings: In this report, we demonstrate efficient gene replacement in a model filamentous fungus, Neurospora crassa, with the CRISPR/Cas9 system. We utilize Cas9 endonuclease and single crRNA:tracrRNA chimeric guide RNA (gRNA) to: (1) replace the endogenous promoter of clr-2 with the ?-tubulin promoter, and (2) introduce a codon optimized fire fly luciferase under the control of the gsy-1 promoter at the csr-1 locus. CLR-2 is one of the core transcription factors that regulate the expression of cellulases, and GSY-1 regulates the conversion of glucose into glycogen. We show that the ?-tubulin promoter driven clr-2 strain shows increased expression of cellulases, and gsy-1-luciferase reporter strain can be easily screened with a bioluminescence assay. Conclusion: CRISPR/Cas9 system works efficiently in Neurospora crassa, which may be adapted to Neurospora natural isolates and other filamentous fungi. It will be beneficial for the filamentous fungal research community to take advantage of CRISPR/Cas9 tool kits that enable genetic perturbations including gene replacement and insertions. Electronic supplementary material: The online version of this article (doi:10.1186/s40694-015-0015-1) contains supplementary material, which is available to authorized users. Twit Text FOAVip Efficient gene editing in Neurospora crassa with CRISPR technology ????Fungal Biol Biotechnol http://www.kidney.de/pget.php?&tw=1&pmid=28955455 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28955455 #FOAvip English Wikipedia <ref>{{Cite pmid|28955455}}</ref> Efficient gene editing in Neurospora crassa with CRISPR technology #MMPMID28955455 Matsu-ura T; Baek M; Kwon J; Hong C Fungal Biol Biotechnol 2015[]; 2 (ä): ä PMID28955455show ga Background: Efficient gene editing is a critical tool for investigating molecular mechanisms of cellular processes and engineering organisms for numerous purposes ranging from biotechnology to medicine. Recently developed RNA-guided CRISPR/Cas9 technology has been used for efficient gene editing in various organisms, but has not been tested in a model filamentous fungus, Neurospora crassa. Findings: In this report, we demonstrate efficient gene replacement in a model filamentous fungus, Neurospora crassa, with the CRISPR/Cas9 system. We utilize Cas9 endonuclease and single crRNA:tracrRNA chimeric guide RNA (gRNA) to: (1) replace the endogenous promoter of clr-2 with the ?-tubulin promoter, and (2) introduce a codon optimized fire fly luciferase under the control of the gsy-1 promoter at the csr-1 locus. CLR-2 is one of the core transcription factors that regulate the expression of cellulases, and GSY-1 regulates the conversion of glucose into glycogen. We show that the ?-tubulin promoter driven clr-2 strain shows increased expression of cellulases, and gsy-1-luciferase reporter strain can be easily screened with a bioluminescence assay. Conclusion: CRISPR/Cas9 system works efficiently in Neurospora crassa, which may be adapted to Neurospora natural isolates and other filamentous fungi. It will be beneficial for the filamentous fungal research community to take advantage of CRISPR/Cas9 tool kits that enable genetic perturbations including gene replacement and insertions. Electronic supplementary material: The online version of this article (doi:10.1186/s40694-015-0015-1) contains supplementary material, which is available to authorized users. äEfficient gene editing in Neurospora crassa with CRISPR technology (epmc).pdf DeepDyve Pubget Overpricing