Targeted insertion of large genetic payloads using cas directed LINE-1 reverse transcriptase

A difficult genome editing goal is the site-specific insertion of large genetic constructs. Here we describe the GENEWRITE system, where site-specific targetable activity of Cas endonucleases is coupled with the reverse transcriptase activity of the ORF2p protein of the human retrotransposon LINE-1. This is accomplished by providing two RNAs: a guide RNA targeting Cas endonuclease activity and an appropriately designed payload RNA encoding the desired insertion. Using E. coli as a simple platform for development and deployment, we show that with proper payload design and co-expression of helper proteins, GENEWRITE can enable insertion of large genetic payloads to precise locations, although with off-target effects, using the described approach. Based upon these results, we describe a potential strategy for implementation of GENEWRITE in more complex systems.

SIFAT KEPRIMAAN MODUL SEDERHANA CHEN UNTUK GRAF 𝐴∞

Let 𝐾𝐾 be a field, 𝐸𝐸 is a directed graph. Let 𝐴𝐴~ is a directed line graph. Suppose that 𝑉𝑉[𝑝𝑝] is a class of Chen simple module for the Leavitt path algebra (𝐿𝐿𝐾𝐾 (𝐸𝐸)), with [p] being equivalent classes containing an infinite path. An infinite path p is an infinite sequence from the sides of a graph. In this paper it will be shown that 𝑉𝑉[𝑝𝑝]is not a prime module of the Leavitt path algebra for graph 𝐴𝐴∞ .Keywords : Leavitt path algebra, Graph 𝐴𝐴~, Chen simple modules, Prime modules

AN EXTRAPOLATORY QUADRATURE RULEFOR ANALYTIC FUNCTIONS

A quadrature rule for the numerical evaluation of integrals of analytic functions along directed line segments in the complex plane has been formulated using the transformed rule based on Gauss Legendre two point quadrature formulas and an interpolatory three point rule. The degree of precision has been increased from five to seven