scholarly journals Nuclease Target Site Selection for Maximizing On-target Activity and Minimizing Off-target Effects in Genome Editing

2016 ◽  
Vol 24 (3) ◽  
pp. 475-487 ◽  
Author(s):  
Ciaran M Lee ◽  
Thomas J Cradick ◽  
Eli J Fine ◽  
Gang Bao
Keyword(s):  
Genome Editing ◽  
Site Selection ◽  
Target Site ◽  
Target Site Selection ◽  
Selection For ◽  
Target Activity ◽  
Target Effects

Mapping target site selection for the non-specific nuclease activities of retroviral integrase

2000 ◽  
Vol 66 (1) ◽  
pp. 87-100 ◽  
Author(s):  
Michael Katzman ◽  
Malgorzata Sudol ◽  
Jeffrey S. Pufnock ◽  
Shawn Zeto ◽  
Lynn M. Skinner
Keyword(s):  
Site Selection ◽  
Target Site ◽  
Target Site Selection ◽  
Selection For ◽  
Retroviral Integrase

Target site selection for an RNA-cleaving catalytic DNA

10.1038/8658 ◽  
1999 ◽  
Vol 17 (5) ◽  
pp. 480-486 ◽  
Author(s):  
Murray J. Cairns ◽  
Toni M. Hopkins ◽  
Craig Witherington ◽  
Li Wang ◽  
Lun-Quan Sun
Keyword(s):  
Site Selection ◽  
Target Site ◽  
Target Site Selection ◽  
Selection For ◽  
Catalytic Dna

scholarly journals Target Site Selection by Tn7:attTn7 Transcription and Target Activity

2000 ◽  
Vol 182 (11) ◽  
pp. 3310-3313 ◽  
Author(s):  
Robert T. DeBoy ◽  
Nancy L. Craig
Keyword(s):  
Host Cell ◽  
Site Selection ◽  
High Frequency ◽  
Cell Metabolism ◽  
Target Site ◽  
Specific Site ◽  
Target Site Selection ◽  
Target Activity ◽  
Bacterial Transposon

ABSTRACT The bacterial transposon Tn7 inserts at high frequency into a specific site called attTn7, which is present in the chromosomes of many bacteria. We show here that transcription of a nearby gene, glmS, decreases the frequency of Tn7 insertion intoattTn7, thus providing a link between Tn7 transposition and host cell metabolism.

Ezrin mRNA target site selection for DNAzymes using secondary structure and hybridization thermodynamics

Tumor Biology ◽  
2011 ◽  
Vol 32 (4) ◽  
pp. 809-817 ◽  
Author(s):  
YaoFei Wang ◽  
JingNan Shen ◽  
XiFu Shang ◽  
Jin Wang ◽  
JingChun Li ◽  
...  
Keyword(s):  
Secondary Structure ◽  
Site Selection ◽  
Target Site ◽  
Target Site Selection ◽  
Selection For

scholarly journals Cas12a is a dynamic and precise RNA-guided nuclease without off-target activity on λ-DNA

2021 ◽  
Author(s):  
Bijoya Paul ◽  
Loic Chaubet ◽  
Emma Verver ◽  
Guillermo Montoya
Keyword(s):  
Dna Binding ◽  
Genome Editing ◽  
Optical Tweezers ◽  
Target Site ◽  
Target Dna ◽  
Multiple Binding ◽  
Target Sites ◽  
Dna Binding And Cleavage ◽  
Target Activity ◽  
Insight Into

Cas12a is an RNA-guided endonuclease that is emerging as a powerful genome-editing tool. Here we combined optical tweezers with fluorescence to monitor Cas12a binding onto λ-DNA, providing insight into its DNA binding and cleavage mechanisms. At low forces Cas12a binds DNA specifically with two off-target sites, while at higher forces numerous binding events appear driven by the mechanical distortion of the DNA and partial matches to the crRNA. Despite the multiple binding events, cleavage is only observed on the target site at low forces, when the DNA is flexible. Activity assays show that the preferential off-target sites are not cleaved, and the λ-DNA is severed at the target site. This precision is also observed in Cas12a variants where the specific dsDNA and the unspecific ssDNA cleavage are dissociated or nick the target DNA. We propose that Cas12a and its variants are precise endonucleases that efficiently scan the DNA for its target but only cleave the selected site in the λ-DNA.

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