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10.1038/s41467-020-18615-1 http://scihub22266oqcxt.onion/10.1038/s41467-020-18615-1 32999292!7528031!32999292     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Nat+Commun 2020 ; 11 (1): 4906 Nephropedia Template TP {{tp|p=32999292|t=2020. Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection |pdf=|usr=}}{{32999292}} gab.com Text Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection JO: Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=32999292 #FOAvip The CRISPR-Cas12a RNA-guided complexes have tremendous potential for nucleic acid detection but are limited to the picomolar detection limit without an amplification step. Here, we develop a platform with engineered crRNAs and optimized conditions that enabled us to detect various clinically relevant nucleic acid targets with higher sensitivity, achieving a limit of detection in the femtomolar range without any target pre-amplification step. By extending the 3'- or 5'-ends of the crRNA with different lengths of ssDNA, ssRNA, and phosphorothioate ssDNA, we discover a self-catalytic behavior and an augmented rate of LbCas12a-mediated collateral cleavage activity as high as 3.5-fold compared to the wild-type crRNA and with significant improvement in specificity for target recognition. Particularly, the 7-mer DNA extension to crRNA is determined to be universal and spacer-independent for enhancing the sensitivity and specificity of LbCas12a-mediated nucleic acid detection. We perform a detailed characterization of our engineered ENHANCE system with various crRNA modifications, target types, reporters, and divalent cations. With isothermal amplification of SARS-CoV-2 RNA using RT-LAMP, the modified crRNAs are incorporated in a paper-based lateral flow assay that can detect the target with up to 23-fold higher sensitivity within 40-60 min. Twit Text FOAVip Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection ????Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=32999292 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32999292 #FOAvip English Wikipedia <ref>{{Cite pmid|32999292}}</ref> Enhancement of trans-cleavage activity of Cas12a with engineered crRNA enables amplified nucleic acid detection #MMPMID32999292 Nguyen LT; Smith BM; Jain PK Nat Commun 2020[Sep]; 11 (1): 4906 PMID32999292show ga The CRISPR-Cas12a RNA-guided complexes have tremendous potential for nucleic acid detection but are limited to the picomolar detection limit without an amplification step. Here, we develop a platform with engineered crRNAs and optimized conditions that enabled us to detect various clinically relevant nucleic acid targets with higher sensitivity, achieving a limit of detection in the femtomolar range without any target pre-amplification step. By extending the 3'- or 5'-ends of the crRNA with different lengths of ssDNA, ssRNA, and phosphorothioate ssDNA, we discover a self-catalytic behavior and an augmented rate of LbCas12a-mediated collateral cleavage activity as high as 3.5-fold compared to the wild-type crRNA and with significant improvement in specificity for target recognition. Particularly, the 7-mer DNA extension to crRNA is determined to be universal and spacer-independent for enhancing the sensitivity and specificity of LbCas12a-mediated nucleic acid detection. We perform a detailed characterization of our engineered ENHANCE system with various crRNA modifications, target types, reporters, and divalent cations. With isothermal amplification of SARS-CoV-2 RNA using RT-LAMP, the modified crRNAs are incorporated in a paper-based lateral flow assay that can detect the target with up to 23-fold higher sensitivity within 40-60 min. |Bacterial Proteins/*metabolism[MESH] |Betacoronavirus/*genetics/isolation & purification[MESH] |COVID-19[MESH] |COVID-19 Testing[MESH] |CRISPR-Associated Proteins/*metabolism[MESH] |CRISPR-Cas Systems[MESH] |Clinical Laboratory Techniques[MESH] |Clustered Regularly Interspaced Short Palindromic Repeats[MESH] |Coronavirus Infections/diagnosis/virology[MESH] |DNA, Single-Stranded[MESH] |Endodeoxyribonucleases/*metabolism[MESH] |Nucleic Acid Amplification Techniques/*methods[MESH] |Pandemics[MESH] |Pneumonia, Viral[MESH] |RNA, Guide, CRISPR-Cas Systems/genetics[MESH] |RNA, Viral/genetics/*isolation & purification[MESH] |SARS-CoV-2[MESH]Enhancement of trans-cleavage activity of Cas12a with engineered crRNA(epmc).pdf DeepDyve Pubget Overpricing