Ryan Thomas Leenay

Works (7)

Updated: August 26th, 2023 21:16

2020 journal article

A positive, growth-based PAM screen identifies noncanonical motifs recognized by the S. pyogenes Cas9

SCIENCE ADVANCES, 6(29).

By: D. Collias n, R. Leenay n, R. Slotkowski n, Z. Zuo *, S. Collins n, B. McGirr n, J. Liu *, C. Beisel n

TL;DR: It is reported that SpyCas9 also recognizes sequences within an N(A/C/T)GG motif, which could have implications for precise editing, off-target predictions, and CRISPR-based immunity. (via Semantic Scholar)
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Source: Web Of Science
Added: August 10, 2020

2019 journal article

Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis

NATURE COMMUNICATIONS, 10.

By: C. Liao  n, F. Ttofali n, R. Slotkowski n, S. Denny n, T. Cecil n, R. Leenay n, A. Keung n , C. Beisel*

MeSH headings : Base Sequence; CRISPR-Associated Proteins / metabolism; Clustered Regularly Interspaced Short Palindromic Repeats / genetics; DNA / genetics; Endonucleases / metabolism; Gene Library; Genetic Techniques; HEK293 Cells; Humans; Nucleic Acid Conformation; Plasmids / metabolism; RNA / biosynthesis; RNA, Messenger / genetics; RNA, Messenger / metabolism; Saccharomyces cerevisiae / metabolism
TL;DR: A modular, one-pot scheme called CRATES to construct CRISPR arrays and array libraries that can facilitate diverse multiplexing applications and help identify factors impacting crRNA biogenesis is presented. (via Semantic Scholar)
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Sources: Web Of Science, NC State University Libraries
Added: July 22, 2019

2018 journal article

Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods

BIOTECHNOLOGY JOURNAL, 14(3).

By: R. Leenay*, J. Vento*, M. Shah*, M. Martino *, F. Leulier * & C. Beisel*

author keywords: genome editing; lactobacilli; recombineering; shuttle plasmids
MeSH headings : CRISPR-Cas Systems / genetics; Clustered Regularly Interspaced Short Palindromic Repeats / genetics; Gene Editing / methods; Lactobacillus plantarum / genetics; Plasmids / genetics; Recombinases / genetics; Recombination, Genetic / genetics
TL;DR: Surprising differences for Cas9-mediated genome editing between methods and related strains are highlighted, arguing for the need for multiple, distinct methods when performing CRISPR-based editing in bacteria. (via Semantic Scholar)
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Source: Web Of Science
Added: March 18, 2019

2016 review

Deciphering, Communicating, and Engineering the CRISPR PAM

[Review of ]. JOURNAL OF MOLECULAR BIOLOGY, 429(2), 177–191.

By: R. Leenay n & C. Beisel n

author keywords: Cas9; Cpf1; CRISPR-Cas systems; PFS; rPAM
MeSH headings : Amino Acid Motifs; CRISPR-Cas Systems; Clustered Regularly Interspaced Short Palindromic Repeats / genetics; DNA, Bacterial / genetics; Gene Library; Genetic Loci; Plasmids / genetics; Protein Engineering; RNA / genetics; RNA / metabolism; Staphylococcus epidermidis / genetics; Streptococcus thermophilus / genetics
TL;DR: The properties of the CRISPR PAM and the emerging tools for determining, visualizing, and engineering PAM recognition are discussed and a standard means of orienting the PAM is proposed to simplify how its location and sequence are communicated. (via Semantic Scholar)
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Added: August 6, 2018

2016 journal article

Identifying and Visualizing Functional PAM Diversity across CRISPR-Cas Systems

MOLECULAR CELL, 62(1), 137–147.

By: R. Leenay n, K. Maksimchuk n, R. Slotkowski n, R. Agrawal n, A. Gomaa n, A. Briner n, R. Barrangou n , C. Beisel n

MeSH headings : Bacillus / chemistry; Bacillus / metabolism; CRISPR-Associated Proteins / chemistry; CRISPR-Associated Proteins / genetics; CRISPR-Cas Systems; Escherichia coli / chemistry; Escherichia coli / metabolism; Francisella / chemistry; Francisella / metabolism; High-Throughput Screening Assays / methods; Protein Structure, Tertiary; Streptococcus thermophilus / chemistry; Streptococcus thermophilus / metabolism
TL;DR: An in vivo, positive, and tunable screen termed PAM-SCANR (PAM screen achieved by NOT-gate repression) to elucidate functional PAMs as well as an interactive visualization scheme termed the PAM wheel to convey individual PAM sequences and their activities to accelerate ability to understand and exploit the multitude of CRISPR-Cas systems in nature. (via Semantic Scholar)
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Sources: Web Of Science, ORCID, NC State University Libraries
Added: August 6, 2018

2015 review

Current and future prospects for CRISPR-based tools in bacteria

[Review of ]. BIOTECHNOLOGY AND BIOENGINEERING, 113(5), 930–943.

By: M. Luo n, R. Leenay n & C. Beisel n

author keywords: antimicrobials; Cas9; genetic control; genetic circuits; genome engineering; undomesticated microbes
MeSH headings : Anti-Bacterial Agents / pharmacology; Bacteria / drug effects; Bacteria / genetics; Bacterial Infections / drug therapy; CRISPR-Cas Systems; Clustered Regularly Interspaced Short Palindromic Repeats; Drug Discovery; Drug Resistance, Bacterial; Gene Editing / methods; Gene Expression Regulation, Bacterial; Genetic Engineering / methods; Genome, Bacterial; Humans
TL;DR: Overall, CRISPR‐Cas systems have formed the basis of an ever‐expanding genetic toolbox and hold tremendous potential for the future understanding and engineering of the bacterial world. (via Semantic Scholar)
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Source: Web Of Science
Added: August 6, 2018

2014 journal article

Repurposing endogenous type I CRISPR-Cas systems for programmable gene repression

NUCLEIC ACIDS RESEARCH, 43(1), 674–681.

By: M. Luo n, A. Mullis n, R. Leenay n & C. Beisel n

MeSH headings : CRISPR-Associated Proteins / genetics; CRISPR-Cas Systems; Cell Engineering; Escherichia coli / genetics; Gene Deletion; Transcription, Genetic
TL;DR: It is found that deletion of the signature cas3 gene converted this immune system in Escherichia coli into a programmable gene regulator capable of reversible gene silencing of heterologous and endogenous genes. (via Semantic Scholar)
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Source: Web Of Science
Added: August 6, 2018

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