CRISPR/Cas9 Mutagenesis of UL21 in Multiple Strains of Herpes Simplex Virus Reveals Differential Requirements for pUL21 in Viral Replication

Viruses
Renée L Finnen, Bruce W Banfield

Abstract

Studies from multiple laboratories using different strains or species of herpes simplex virus (HSV) with deletions in UL21 have yielded conflicting results regarding the necessity of pUL21 in HSV infection. To resolve this discrepancy, we utilized CRISPR/Cas9 mutagenesis to isolate pUL21 deficient viruses in multiple HSV backgrounds, and performed a side-by-side comparison of the cell-to-cell spread and replication phenotypes of these viruses. These analyses confirmed previous studies implicating the involvement of pUL21 in cell-to-cell spread of HSV. Cell-to-cell spread of HSV-2 was more greatly affected by the lack of pUL21 than HSV-1, and strain-specific differences in the requirement for pUL21 in cell-to-cell spread were also noted. HSV-2 strain 186 lacking pUL21 was particularly crippled in both cell-to-cell spread and viral replication in non-complementing cells, in comparison to other HSV strains lacking pUL21, suggesting that the strict requirement for pUL21 by strain 186 may not be representative of the HSV-2 species as a whole. This work highlights CRISPR/Cas9 technology as a useful tool for rapidly constructing deletion mutants of alphaherpesviruses, regardless of background strain, and should find great utility when...Continue Reading

References

Apr 1, 1973·Virology·F L Graham, A J van der Eb
Aug 5, 1999·Methods in Enzymology·B C HorsburghF Tufaro
Dec 6, 2001·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·H TakakuwaY Nishiyama
May 2, 2006·Journal of Virological Methods·William W GieraschDavid A Leib
Apr 21, 2009·Methods in Molecular Biology·Bruce W Banfield, Gregory A Bird
Jan 1, 2010·Journal of Virology·Amy L HarperJohn W Wills
Nov 15, 2012·Proceedings of the National Academy of Sciences of the United States of America·Jun HanJohn W Wills
Jan 5, 2013·Science·Le CongFeng Zhang
Mar 15, 2013·Journal of Virology·Valerie Le SageBruce W Banfield
Mar 29, 2014·Methods in Molecular Biology·Nancy M Sawtell, Richard L Thompson
Dec 30, 2014·Journal of Virology·Claire M MetrickEkaterina E Heldwein

❮ Previous
Next ❯

Citations

Apr 1, 2020·PLoS Pathogens·Julia SehlThomas C Mettenleiter
Sep 28, 2018·Reviews in Medical Virology·Saeedeh EbrahimiMaryam Tabasi
Dec 18, 2020·Frontiers in Cellular and Infection Microbiology·Julianne VilelaMuhammad Munir

❮ Previous
Next ❯

Methods Mentioned

BETA
glycosylation
PCR
scraping
electrophoresis

Software Mentioned

Pro
Image

Related Concepts

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

Virology & CRISPR

This feed focuses on the virology of CRISPR and its use in developing CRISPR-Cas systems. Discover the latest research here.