Can In-Fusion be used to clone a microRNA (miRNA) precursor

-Sequences for microRNA precursors and flanking genomic DNA can be obtained from a number of public databases, including GenBank and EMBL-Bank. The UCSC Genomic Bioniformatics Site hosts an easy-to-navigate genomic database which tracks miRNAs. The Sanger Institute hosts miRBase, a compilation of known miRNA sequences.
-100–300 bp of DNA flanking the miRNA precursor is amplified from genomic DNA for cloning into the 3' UTR of a fluorescent protein, carried by an miRNA expression vector. The flanking DNA ensures efficient processing by Drosha.
-For In-Fusion Cloning, the miRNA precursor (100–300 bp) is PCR amplified, incorporating 15-bp overhangs homologous to the termini of the miRNA expression vector. The vector should be linearized at the fluorescent protein's 3' UTR. The suggested miRNA precursor-to-vector molar ratio for the cloning reaction is 3–5:1, depending on the precursor length. Optimal molar ratios must be determined empirically.
-In the cell, the miRNA precursor is coexpressed with the fluorescent protein (as described in Figure 2 of this tech note), allowing both of the following:
*Expression of the fluorescent protein, resulting in fluorescent-cell labeling.
*miRNA precursor processing, resulting in targeted gene knockdown in fluorescently labeled cells.
-We offer Tet-inducible miRNA expression systems and vectors with either a red or green fluorescent protein marker.

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