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A study of E. coli RNA polymerase‐Template interaction utilizing CRISPR/dCas9 protein
Author(s) -
King Maxwell J.M.,
Rohlman Christopher E.
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.07125
Subject(s) - transcription (linguistics) , rna polymerase , rna dependent rna polymerase , polymerase , rna , biology , rna polymerase ii , dna , microbiology and biotechnology , genetics , gene , gene expression , promoter , philosophy , linguistics
This project focuses on sequence‐specific E. coli RNA polymerase transcription pausing using dCas9 protein. dCas9 is one type of Cas9 protein which is an RNA‐guided enzyme that cleaves DNA at specific locations. dCas9 lacks the ability to cut DNA or RNA. However, this dead‐Cas9 (dCas9) retains its ability to bind DNA at a specific location when guided by a complementary RNA molecule. In order to bind to desired location of DNA, dCas9 needs this small RNA that can guide it to bind at the targeted DNA sequence. When dCas9 binds to the DNA sequence, it blocks transcription, which is the process of copying DNA into RNA with the help of enzyme called RNA polymerase. The goal of this project is to investigate the structural and chemical interaction between dCas9, DNA and E. coli RNA polymerase at the point of the transcription blockade. A second goal of this project is to study the behavior of the E. coli RNA polymerase following a targeted pause in the transcription process